Nucleic acid and corresponding protein entitled 98P4B6 useful in treatment and detection of cancer

ABSTRACT

A novel gene 098P4B6 (also designated STEAP-2) and its encoded protein, and variants thereof, are described wherein 98P4B6 exhibits tissue specific expression in normal adult tissue, and is aberrantly expressed in the cancers listed in Table I. Consequently, 98P4B6 provides a diagnostic, prognostic, prophylactic and/or therapeutic target for cancer. The 98P4B6 gene or fragment thereof, or its encoded protein, or variants thereof, or a fragment thereof, can be used to elicit a humoral or cellular immune response; antibodies or T cells reactive with 98P4B6 can be used in active or passive immunization.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No. 11/068,859, filed Feb. 28, 2005, now abandoned, which is a continuation of U.S. patent application Ser. No. 10/862,182, filed Jun. 4, 2004, now abandoned, which is a continuation of U.S. patent application Ser. No. 10/455,822, filed Jun. 4, 2003, now abandoned, which claims the benefit of U.S. Provisional Application No. 60/435,480, filed Dec. 20, 2002, and which is a continuation-in-part of U.S. patent application Ser. No. 10/407,484, filed Apr. 4, 2003, now abandoned, which is a continuation-in-part of U.S. patent application Ser. No. 09/455,486, filed Dec. 6, 1999, now issued as U.S. Pat. No. 6,833,438 on Dec. 21, 2004, which is a continuation-in-part of U.S. patent application Ser. No. 09/323,873, filed Jun. 1, 1999, now issued as U.S. Pat. No. 6,329,503 on Dec. 11, 2001, which claims the benefit of U.S. Provisional Application No. 60/091,183, filed Jun. 30, 1998 and U.S. Provisional Application No. 60/087,520, filed Jun. 1, 1998. This application relates to U.S. Provisional Application No. 60/296,656, filed Jun. 6, 2001, U.S. patent application Ser. No. 10/165,044, filed Jun. 6, 2002, and U.S. patent application Ser. No. 10/753,195, filed Jan. 6, 2004. The contents of the applications listed in this paragraph are fully incorporated by reference herein.

STATEMENT OF RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH

Not applicable.

FIELD OF THE INVENTION

The invention described herein relates to genes and their encoded proteins, termed 98P4B6 or STEAP-2, expressed in certain cancers, and to diagnostic and therapeutic methods and compositions useful in the management of cancers that express 98P4B6.

BACKGROUND OF THE INVENTION

Cancer is the second leading cause of human death next to coronary disease. Worldwide, millions of people die from cancer every year. In the United States alone, as reported by the American Cancer Society, cancer causes the death of well over a half-million people annually, with over 1.2 million new cases diagnosed per year. While deaths from heart disease have been declining significantly, those resulting from cancer generally are on the rise. In the early part of the next century, cancer is predicted to become the leading cause of death.

Worldwide, several cancers stand out as the leading killers. In particular, carcinomas of the lung, prostate, breast, colon, pancreas, and ovary represent the primary causes of cancer death. These and virtually all other carcinomas share a common lethal feature. With very few exceptions, metastatic disease from a carcinoma is fatal. Moreover, even for those cancer patients who initially survive their primary cancers, common experience has shown that their lives are dramatically altered. Many cancer patients experience strong anxieties driven by the awareness of the potential for recurrence or treatment failure. Many cancer patients experience physical debilitations following treatment. Furthermore, many cancer patients experience a recurrence.

Worldwide, prostate cancer is the fourth most prevalent cancer in men. In North America and Northern Europe, it is by far the most common cancer in males and is the second leading cause of cancer death in men. In the United States alone, well over 30,000 men die annually of this disease—second only to lung cancer. Despite the magnitude of these figures, there is still no effective treatment for metastatic prostate cancer. Surgical prostatectomy, radiation therapy, hormone ablation therapy, surgical castration and chemotherapy continue to be the main treatment modalities. Unfortunately, these treatments are ineffective for many and are often associated with undesirable consequences.

On the diagnostic front, the lack of a prostate tumor marker that can accurately detect early-stage, localized tumors remains a significant limitation in the diagnosis and management of this disease. Although the serum prostate specific antigen (PSA) assay has been a very useful tool, however its specificity and general utility is widely regarded as lacking in several important respects.

Progress in identifying additional specific markers for prostate cancer has been improved by the generation of prostate cancer xenografts that can recapitulate different stages of the disease in mice. The LAPC (Los Angeles Prostate Cancer) xenografts are prostate cancer xenografts that have survived passage in severe combined immune deficient (SCID) mice and have exhibited the capacity to mimic the transition from androgen dependence to androgen independence (Klein et al., 1997, Nat. Med. 3:402). More recently identified prostate cancer markers include PCTA-1 (Su et al., 1996, Proc. Natl. Acad. Sci. USA 93: 7252), prostate-specific membrane (PSM) antigen (Pinto et al., Clin Cancer Res 1996 Sep. 2 (9): 1445-51), STEAP (Hubert, et al., Proc Natl Acad Sci USA. 1999 Dec. 7; 96(25): 14523-8) and prostate stem cell antigen (PSCA) (Reiter et al., 1998, Proc. Natl. Acad. Sci. USA 95: 1735).

While previously identified markers such as PSA, PSM, PCTA and PSCA have facilitated efforts to diagnose and treat prostate cancer, there is need for the identification of additional markers and therapeutic targets for prostate and related cancers in order to further improve diagnosis and therapy.

Renal cell carcinoma (RCC) accounts for approximately 3 percent of adult malignancies. Once adenomas reach a diameter of 2 to 3 cm, malignant potential exists. In the adult, the two principal malignant renal tumors are renal cell adenocarcinoma and transitional cell carcinoma of the renal pelvis or ureter. The incidence of renal cell adenocarcinoma is estimated at more than 29,000 cases in the United States, and more than 11,600 patients died of this disease in 1998. Transitional cell carcinoma is less frequent, with an incidence of approximately 500 cases per year in the United States.

Surgery has been the primary therapy for renal cell adenocarcinoma for many decades. Until recently, metastatic disease has been refractory to any systemic therapy. With recent developments in systemic therapies, particularly immunotherapies, metastatic renal cell carcinoma may be approached aggressively in appropriate patients with a possibility of durable responses. Nevertheless, there is a remaining need for effective therapies for these patients.

Of all new cases of cancer in the United States, bladder cancer represents approximately 5 percent in men (fifth most common neoplasm) and 3 percent in women (eighth most common neoplasm). The incidence is increasing slowly, concurrent with an increasing older population. In 1998, there was an estimated 54,500 cases, including 39,500 in men and 15,000 in women. The age-adjusted incidence in the United States is 32 per 100,000 for men and eight per 100,000 in women. The historic male/female ratio of 3:1 may be decreasing related to smoking patterns in women. There were an estimated 11,000 deaths from bladder cancer in 1998 (7,800 in men and 3,900 in women). Bladder cancer incidence and mortality strongly increase with age and will be an increasing problem as the population becomes more elderly.

Most bladder cancers recur in the bladder. Bladder cancer is managed with a combination of transurethral resection of the bladder (TUR) and intravesical chemotherapy or immunotherapy. The multifocal and recurrent nature of bladder cancer points out the limitations of TUR. Most muscle-invasive cancers are not cured by TUR alone. Radical cystectomy and urinary diversion is the most effective means to eliminate the cancer but carry an undeniable impact on urinary and sexual function. There continues to be a significant need for treatment modalities that are beneficial for bladder cancer patients.

An estimated 130,200 cases of colorectal cancer occurred in 2000 in the United States, including 93,800 cases of colon cancer and 36,400 of rectal cancer. Colorectal cancers are the third most common cancers in men and women. Incidence rates declined significantly during 1992-1996 (−2.1% per year). Research suggests that these declines have been due to increased screening and polyp removal, preventing progression of polyps to invasive cancers. There were an estimated 56,300 deaths (47,700 from colon cancer, 8,600 from rectal cancer) in 2000, accounting for about 11% of all U.S. cancer deaths.

At present, surgery is the most common form of therapy for colorectal cancer, and for cancers that have not spread, it is frequently curative. Chemotherapy, or chemotherapy plus radiation, is given before or after surgery to most patients whose cancer has deeply perforated the bowel wall or has spread to the lymph nodes. A permanent colostomy (creation of an abdominal opening for elimination of body wastes) is occasionally needed for colon cancer and is infrequently required for rectal cancer. There continues to be a need for effective diagnostic and treatment modalities for colorectal cancer.

There were an estimated 164,100 new cases of lung and bronchial cancer in 2000, accounting for 14% of all U.S. cancer diagnoses. The incidence rate of lung and bronchial cancer is declining significantly in men, from a high of 86.5 per 100,000 in 1984 to 70.0 in 1996. In the 1990s, the rate of increase among women began to slow. In 1996, the incidence rate in women was 42.3 per 100,000.

Lung and bronchial cancer caused an estimated 156,900 deaths in 2000, accounting for 28% of all cancer deaths. During 1992-1996, mortality from lung cancer declined significantly among men (−1.7% per year) while rates for women were still significantly increasing (0.9% per year). Since 1987, more women have died each year of lung cancer than breast cancer, which, for over 40 years, was the major cause of cancer death in women. Decreasing lung cancer incidence and mortality rates most likely resulted from decreased smoking rates over the previous 30 years; however, decreasing smoking patterns among women lag behind those of men. Of concern, although the declines in adult tobacco use have slowed, tobacco use in youth is increasing again.

Treatment options for lung and bronchial cancer are determined by the type and stage of the cancer and include surgery, radiation therapy, and chemotherapy. For many localized cancers, surgery is usually the treatment of choice. Because the disease has usually spread by the time it is discovered, radiation therapy and chemotherapy are often needed in combination with surgery. Chemotherapy alone or combined with radiation is the treatment of choice for small cell lung cancer; on this regimen, a large percentage of patients experience remission, which in some cases is long lasting. There is however, an ongoing need for effective treatment and diagnostic approaches for lung and bronchial cancers.

An estimated 182,800 new invasive cases of breast cancer were expected to occur among women in the United States during 2000. Additionally, about 1,400 new cases of breast cancer were expected to be diagnosed in men in 2000. After increasing about 4% per year in the 1980s, breast cancer incidence rates in women have leveled off in the 1990s to about 110.6 cases per 100,000.

In the U.S. alone, there were an estimated 41,200 deaths (40,800 women, 400 men) in 2000 due to breast cancer. Breast cancer ranks second among cancer deaths in women. According to the most recent data, mortality rates declined significantly during 1992-1996 with the largest decreases in younger women, both white and black. These decreases were probably the result of earlier detection and improved treatment.

Taking into account the medical circumstances and the patient's preferences, treatment of breast cancer may involve lumpectomy (local removal of the tumor) and removal of the lymph nodes under the arm; mastectomy (surgical removal of the breast) and removal of the lymph nodes under the arm; radiation therapy; chemotherapy; or hormone therapy. Often, two or more methods are used in combination. Numerous studies have shown that, for early stage disease, long-term survival rates after lumpectomy plus radiotherapy are similar to survival rates after modified radical mastectomy. Significant advances in reconstruction techniques provide several options for breast reconstruction after mastectomy. Recently, such reconstruction has been done at the same time as the mastectomy.

Local excision of ductal carcinoma in situ (DCIS) with adequate amounts of surrounding normal breast tissue may prevent the local recurrence of the DCIS. Radiation to the breast and/or tamoxifen may reduce the chance of OCIS occurring in the remaining breast tissue. This is important because DCIS, if left untreated, may develop into invasive breast cancer. Nevertheless, there are serious side effects or sequelae to these treatments. There is, therefore, a need for efficacious breast cancer treatments.

There were an estimated 23,100 new cases of ovarian cancer in the United States in 2000. It accounts for 4% of all cancers among women and ranks second among gynecologic cancers. During 1992-1996, ovarian cancer incidence rates were significantly declining. Consequent to ovarian cancer, there were an estimated 14,000 deaths in 2000. Ovarian cancer causes more deaths than any other cancer of the female reproductive system.

Surgery, radiation therapy, and chemotherapy are treatment options for ovarian cancer. Surgery usually includes the removal of one or both ovaries, the fallopian tubes (salpingo-oophorectomy), and the uterus (hysterectomy). In some very early tumors, only the involved ovary will be removed, especially in young women who wish to have children. In advanced disease, an attempt is made to remove all intra-abdominal disease to enhance the effect of chemotherapy. There continues to be an important need for effective treatment options for ovarian cancer.

There were an estimated 28,300 new cases of pancreatic cancer in the United States in 2000. Over the past 20 years, rates of pancreatic cancer have declined in men. Rates among women have remained approximately constant but may be beginning to decline. Pancreatic cancer caused an estimated 28,200 deaths in 2000 in the United States. Over the past 20 years, there has been a slight but significant decrease in mortality rates among men (about −0.9% per year) while rates have increased slightly among women.

Surgery, radiation therapy, and chemotherapy are treatment options for pancreatic cancer. These treatment options can extend survival and/or relieve symptoms in many patients but are not likely to produce a cure for most. There is a significant need for additional therapeutic and diagnostic options for pancreatic cancer.

SUMMARY OF THE INVENTION

The present invention relates to a gene, designated 98P4B6, that has now been found to be over-expressed in the cancer(s) listed in Table I. Northern blot expression analysis of 98P4B6 gene expression in normal tissues shows a restricted expression pattern in adult tissues. The nucleotide (FIG. 2) and amino acid (FIG. 2, and FIG. 3) sequences of 98P4B6 are provided. The tissue-related profile of 98P4B6 in normal adult tissues, combined with the over-expression observed in the tissues listed in Table I, shows that 98P4B6 is aberrantly over-expressed in at least some cancers, and thus serves as a useful diagnostic, prophylactic, prognostic, and/or therapeutic target for cancers of the tissue(s) such as those listed in Table I.

The invention provides polynucleotides corresponding or complementary to all or part of the 98P4B6 genes, mRNAs, and/or coding sequences, preferably in isolated form, including polynucleotides encoding 98P4B6-related proteins and fragments of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more than 25 contiguous amino acids; at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100 or more than 100 contiguous amino acids of a 98P4B6-related protein, as well as the peptides/proteins themselves; DNA, RNA, DNA/RNA hybrids, and related molecules, polynucleotides or oligonucleotides complementary or having at least a 90% homology to the 98P4B6 genes or mRNA sequences or parts thereof, and polynucleotides or oligonucleotides that hybridize to the 98P4B6 genes, mRNAs, or to 98P4B6-encoding polynucleotides. Also provided are means for isolating cDNAs and the genes encoding 98P4B6. Recombinant DNA molecules containing 98P4B6 polynucleotides, cells transformed or transduced with such molecules, and host-vector systems for the expression of 98P4B6 gene products are also provided. The invention further provides antibodies that bind to 98P4B6 proteins and polypeptide fragments thereof, including polyclonal and monoclonal antibodies, murine and other mammalian antibodies, chimeric antibodies, humanized and fully human antibodies, and antibodies labeled with a detectable marker or therapeutic agent. In certain embodiments, there is a proviso that the entire nucleic acid sequence of FIG. 2 is not encoded and/or the entire amino acid sequence of FIG. 2 is not prepared. In certain embodiments, the entire nucleic acid sequence of FIG. 2 is encoded and/or the entire amino acid sequence of FIG. 2 is prepared, either of which are in respective human unit dose forms.

The invention further provides methods for detecting the presence and status of 98P4B6 polynucleotides and proteins in various biological samples, as well as methods for identifying cells that express 98P4B6. A typical embodiment of this invention provides methods for monitoring 98P4B6 gene products in a tissue or hematology sample having or suspected of having some form of growth dysregulation such as cancer.

The invention further provides various immunogenic or therapeutic compositions and strategies for treating cancers that express 98P4B6 such as cancers of tissues listed in Table I, including therapies aimed at inhibiting the transcription, translation, processing or function of 98P4B6 as well as cancer vaccines. In one aspect, the invention provides compositions, and methods comprising them, for treating a cancer that expresses 98P4B6 in a human subject wherein the composition comprises a carrier suitable for human use and a human unit dose of one or more than one agent that inhibits the production or function of 98P4B6. Preferably, the carrier is a uniquely human carrier. In another aspect of the invention, the agent is a moiety that is immunoreactive with 98P4B6 protein. Non-limiting examples of such moieties include, but are not limited to, antibodies (such as single chain, monoclonal, polyclonal, humanized, chimeric, or human antibodies), functional equivalents thereof (whether naturally occurring or synthetic), and combinations thereof. The antibodies can be conjugated to a diagnostic or therapeutic moiety. In another aspect, the agent is a small molecule as defined herein.

In another aspect, the agent comprises one or more than one peptide which comprises a cytotoxic T lymphocyte (CTL) epitope that binds an HLA class I molecule in a human to elicit a CTL response to 98P4B6 and/or one or more than one peptide which comprises a helper T lymphocyte (HTL) epitope which binds an HLA class II molecule in a human to elicit an HTL response. The peptides of the invention may be on the same or on one or more separate polypeptide molecules. In a further aspect of the invention, the agent comprises one or more than one nucleic acid molecule that expresses one or more than one of the CTL or HTL response stimulating peptides as described above. In yet another aspect of the invention, the one or more than one nucleic acid molecule may express a moiety that is immunologically reactive with 98P4B6 as described above. The one or more than one nucleic acid molecule may also be, or encodes, a molecule that inhibits production of 98P4B6. Non-limiting examples of such molecules include, but are not limited to, those complementary to a nucleotide sequence essential for production of 98P4B6 (e.g. antisense sequences or molecules that form a triple helix with a nucleotide double helix essential for 98P4B6 production) or a ribozyme effective to lyse 98P4B6 mRNA.

Note that to determine the starting position of any peptide set forth in Tables VIII-XXI and XXII to XLIX (collectively HLA Peptide Tables) respective to its parental protein, e.g., variant 1, variant 2, etc., reference is made to three factors: the particular variant, the length of the peptide in an HLA Peptide Table, and the Search Peptides in Table VII. Generally, a unique Search Peptide is used to obtain HLA peptides of a particular for a particular variant. The position of each Search Peptide relative to its respective parent molecule is listed in Table VII. Accordingly, if a Search Peptide begins at position “X”, one must add the value “X−1” to each position in Tables VIII-XXI and XXII to XLIX to obtain the actual position of the HLA peptides in their parental molecule. For example, if a particular Search Peptide begins at position 150 of its parental molecule, one must add 150−1, i.e., 149 to each HLA peptide amino acid position to calculate the position of that amino acid in the parent molecule.

One embodiment of the invention comprises an HLA peptide, that occurs at least twice in Tables VIII-XXI and XXII to XLIX collectively, or an oligonucleotide that encodes the HLA peptide. Another embodiment of the invention comprises an HLA peptide that occurs at least once in Tables VIII-XXI and at least once in tables XXII to XLIX, or an oligonucleotide that encodes the HLA peptide.

Another embodiment of the invention is antibody epitopes, which comprise a peptide regions, or an oligonucleotide encoding the peptide region, that has one two, three, four, or five of the following characteristics:

i) a peptide region of at least 5 amino acids of a particular peptide of FIG. 3, in any whole number increment up to the full length of that protein in FIG. 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Hydrophilicity profile of FIG. 5;

ii) a peptide region of at least 5 amino adds of a particular peptide of FIG. 3, in any whole number increment up to the full length of that protein in FIG. 3, that includes an amino acid position having a value equal to or less than 0.5, 0.4, 0.3, 0.2, 0.1, or having a value equal to 0.0, in the Hydropathicity profile of FIG. 6;

iii) a peptide region of at least 5 amino adds of a particular peptide of FIG. 3, in any whole number increment up to the full length of that protein in FIG. 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Percent Accessible Residues profile of FIG. 7;

iv) a peptide region of at least 5 amino adds of a particular peptide of FIG. 3, in any whole number increment up to the full length of that protein in FIG. 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Average Flexibility profile of FIG. 8; or

v) a peptide region of at least 5 amino acids of a particular peptide of FIG. 3, in any whole number increment up to the full length of that protein in FIG. 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Beta-turn profile of FIG. 9.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. The 98P4B6 SSH sequence of 183 nucleotides.

FIG. 2. A) The cDNA and amino acid sequence of 98P4B6 variant 1 (also called “98P4B6 v.1” or “98P4B6 variant 1”) is shown in FIG. 2A. The start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon.

B) The cDNA and amino acid sequence of 98P4B6 variant 2 (also called “98P4B6 v.2”) is shown in FIG. 2B. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 4-138 including the stop codon.

C) The cDNA and amino acid sequence of 98P4B6 variant 3 (also called “98P4B6 v.3”) is shown in FIG. 2C. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 188-1552 including the stop codon.

D) The cDNA and amino acid sequence of 98P4B6 variant 4 (also called “98P4B6 v.4”) is shown in FIG. 2D. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 318-1682 including the stop codon.

E) The cDNA and amino acid sequence of 98P4B6 variant 5 (also called “98P4B6 v.5”) is shown in FIG. 2E. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 318-1577 including the stop codon.

F) The cDNA and amino acid sequence of 98P4B6 variant 6 (also called “98P4B6 v.6”) is shown in FIG. 2F. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 318-1790 including the stop codon.

G) The cDNA and amino acid sequence of 98P4B6 variant 7 (also called “98P4B6 v.7”) is shown in FIG. 2G. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 295-2025 including the stop codon.

H) The cDNA and amino acid sequence of 98P4B6 variant 8 (also called “98P4B6 v.8”) is shown in FIG. 2H. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 394-1866 including the stop codon.

I) The cDNA and amino acid sequence of 98P4B6 variant 9 (also called “98P4B6 v.9”) is shown in FIG. 2I. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon.

J) The cDNA and amino acid sequence of 98P4B6 variant 10 (also called “98P4B6v.10”) is shown in FIG. 2J. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon.

K) The cDNA and amino acid sequence of 98P4B6 variant 11 (also called “98P486 v.11”) is shown in FIG. 2K. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon.

L) The cDNA and amino acid sequence of 98P4B6 variant 12 (also called “98P4B6 v.12”) is shown in FIG. 2L. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon.

M) The cDNA and amino acid sequence of 98P4B6 variant 13 (also called “98P4B6 v.13”) is shown in FIG. 2M. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon.

N) The cDNA and amino acid sequence of 98P4B6 variant 14 (also called “98P4B6 v.14”) is shown in FIG. 2N. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon.

O) The cDNA and amino acid sequence of 98P4B6 variant 15 (also called “98P4B6 v.15”) is shown in FIG. 2O. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon.

P) The cDNA and amino acid sequence of 98P4B6 variant 16 (also called “98P4B6 v.16”) is shown in FIG. 2P. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon.

Q) The cDNA and amino acid sequence of 98P4B6 variant 17 (also called “98P4B6 v.17”) is shown in FIG. 2Q. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon:

R) The cDNA and amino acid sequence of 98P4B6 variant 18 (also called “98P4B6 v.18”) is shown in FIG. 2R. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 355-1719 including the stop codon.

S) The cDNA and amino acid sequence of 98P4B6 variant 19 (also called “98P4B6 v.19”) is shown in FIG. 2S. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 355-1719 including the stop codon.

T) The cDNA and amino acid sequence of 98P4B6 variant 20 (also called “98P4B6 v.20”) is shown in FIG. 2T. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 295-2025 including the stop codon.

U) The cDNA and amino acid sequence of 98P4B6 variant 21 (also called “98P4B6 v.21”) is shown in FIG. 2U. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 295-2025 including the stop codon.

V) The cDNA and amino acid sequence of 98P4B6 variant 22 (also called “98P4B6 v.22”) is shown in FIG. 2V. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 295-2025 including the stop codon.

W) The cDNA and amino acid sequence of 98P4B6 variant 23 (also called 98P4B6 v.23′) is shown in FIG. 2W. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 295-2025 including the stop codon.

X) The cDNA and amino acid sequence of 98P4B6 variant 24 (also called “98P4B6 v.24”) is shown in FIG. 2X The codon for the start methionine is underlined. The open reading frame extends from nucleic add 295-2025 including the stop codon.

Y) The cDNA and amino acid sequence of 98P4B6 variant 25 (also called “98P4B6 v.25”) is shown in FIG. 2Y. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 394-1866 including the stop codon.

Z) The cDNA and amino acid sequence of 98P4B6 variant 26 (also called “98P4B6 v.26”) is shown in FIG. 2Z. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 394-1866 including the stop codon.

M) The cDNA and amino acid sequence of 98P4B6 variant 27 (also called “98P4B6 v.27”) is shown in FIG. 2AA. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 394-1866 including the stop codon.

AB) The cDNA and amino acid sequence of 98P4B6 variant 28 (also called “98P4B6 v.28”) is shown in FIG. 2AB. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 394-1866 including the stop codon.

AC) The cDNA and amino acid sequence of 98P4B6 variant 29 (also called “98P4B6 v.29”) is shown in FIG. 2AC. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 394-1866 including the stop codon.

AD) The cDNA and amino acid sequence of 98P4B6 variant 30 (also called “98P4B6 v.30”) is shown in FIG. 2AD. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 394-1866 including the stop codon.

AE) The cDNA and amino acid sequence of 98P4B6 variant 31 (also called “98P4B6 v.31”) is shown in FIG. 2AE. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 394-1866 including the stop codon.

AF) The cDNA and amino acid sequence of 98P4B6 variant 32 (also called “98P4B6 v.32”) is shown in FIG. 2AF. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 394-1866 including the stop codon.

AG) The cDNA and amino acid sequence of 98P4B6 variant 33 (also called “98P4B6 v.33”) is shown in FIG. 2AG. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 394-1866 including the stop codon.

AH) The cDNA and amino acid sequence of 98P4B6 variant 34 (also called “98P4B6 v.34”) is shown in FIG. 2AH. The codon for the start methionine is underlined. The open reading frame extends from nudcleic acid 394-1866 including the stop codon.

AI) The cDNA and amino acid sequence of 98P4B6 variant 35 (also called “98P4B6 v.35”) is shown in FIG. 2AI. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 394-1866 including the stop codon.

AJ) The cDNA and amino acid sequence of 98P4B6 variant 36 (also called “98P4B6 v.36”) is shown in FIG. 2AJ. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 394-1866 including the stop codon.

AK) The cDNA and amino acid sequence of 98P4B6 variant 37 (also called “98P4B6 v.37”) is shown in FIG. 2AK. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 394-1866 including the stop codon.

AL) The cDNA and amino acid sequence of 98P4B6 variant 38 (also called “98P4B6 v.38”) is shown in FIG. 2AL. The codon for the start methionine is underlined. The open reading frame extends from nucleic add 394-1866 including the stop codon.

FIG. 3.

A) The amino acid sequence of 98P4B6 v.1 is shown in FIG. 3A; it has 454 amino acids.

B) The amino acid sequence of 98P4B6 v.2 is shown in FIG. 3B; it has 45 amino acids.

C) The amino acid sequence of 98P4B6 v.5 is shown in FIG. 3C; it has 419 amino acids.

D) The amino acid sequence of 98P4B6 v.6 is shown in FIG. 3D; it has 490 amino acids.

E) The amino acid sequence of 98P4B6 v.7 is shown in FIG. 3E; it has 576 amino adds.

F) The amino acid sequence of 98P4B6 v.8 is shown in FIG. 3F; it has 490 amino adds.

G) The amino acid sequence of 98P4B6 v.13 is shown in FIG. 3G; it has 454 amino acids.

H) The amino acid sequence of 98P4B6 v.14 is shown in FIG. 3H; it has 454 amino adds:

I) The amino acid sequence of 98P4B6 v.21 is shown in FIG. 3I; it has 576 amino acids.

J) The amino acid sequence of 98P4B6 v.25 is shown in FIG. 3J; it has 490 amino acids.

As used herein, a reference to 98P4B6 includes all variants thereof, including those shown in FIGS. 2, 3, 10, and 11, unless the context dearly indicates otherwise.

FIG. 4. Comparison of 98P4B6 with known genes: Human STAMP1, human six transmembrane-epithelial antigen of prostate 2 and mouse six transmembrane epithelial antigen of prostate 2. FIG. 4(A) Alignment of 98P4B6 variant 1 to human STAMP1 (gi 15418732). FIG. 4(B) Alignment of 98P4B6 variant 1 with human STEAP2 (gi:23308593). FIG. 4(C) Alignment of 98P4B6 variant 1 with mouse STEAP2 (gi 28501136). FIG. 4(D): Clustal Alignment of the three 98P4B6 variants, depicting that 98P4B6 V1B (SEQ ID NO: 94) contains an additional 62 aa at its N-terminus relative to V1 (SEQ ID NO: 95), and that 98P4B6 V2 (SEQ ID NO: 96) carries a I to T point mutation at aa 225 relative to V1.

FIG. 5. Hydrophilicity amino acid profile of 98P4B6v.1, v.2, v.5, v.6, and v.7 determined by computer algorithm sequence analysis using the method of Hopp and Woods (Hopp T. P., Woods K. R., 1981. Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828) accessed on the Protscale website through the ExPasy molecular biology server.

FIG. 6. Hydropathicity amino acid profile of 98P4B6v.1, v.2, v.5, v.6, and v.7 determined by computer algorithm sequence analysis using the method of Kyte and Doolittle (Kyte J., Doolittle R. F., 1982. J. Mol. Biol. 157:105-132) accessed on the ProtScale website through the ExPasy molecular biology server.

FIG. 7. Percent accessible residues amino acid profile of 98P4B6v.1, v.2, v.5, v.6, and v.7 determined by computer algorithm sequence analysis using the method of Janin (Janin J., 1979 Nature 277:491-492) accessed on the ProtScale website through the ExPasy molecular biology server.

FIG. 8. Average flexibility amino acid profile of 98P4B6v.1, v.2, v.5, v.6, and v.7 determined by computer algorithm sequence analysis using the method of Bhaskaran and Ponnuswamy (Bhaskaran R., and Ponnuswamy P. K., 1988. Int. J. Pept. Protein Res. 32:242-255) accessed on the ProtScale website through the ExPasy molecular biology server.

FIG. 9. Beta-turn amino acid profile of 98P4B6v.1, v.2, v.5, v.6, and v.7 determined by computer algorithm sequence analysis using the method of Deleage and Roux (Deleage, G., Roux B. 1987 Protein Engineering 1:289-294) accessed on the ProtScale website through the ExPasy molecular biology server.

FIG. 10. FIG. 10( a): Schematic alignment of SNP variants of 98P4B6 v.1. Variants 98P4B6 v.9 through v.19 were variants with single nucleotide difference from v.1. Though these SNP variants were shown separately, they could also occur in any combinations and in any transcript variants, as shown in FIG. 12, that contains the bases. SNP in regions of other transcript variants, such as v.2, v.6 and v.8, not common with v.1 were not shown here. Numbers correspond to those of 98P4B6 v.1. Black box shows the same sequence as 98P4B6 v.1. SNPs are indicated above the box. FIG. 10( b): Schematic alignment of SNP variants of 98P4B6 v.7. Variants 98P4B6 v.20 through v.24 were variants with single nucleotide difference from v.7. Though these SNP variants were shown separately, they could also occur in any combinations and in any transcript variants, as shown in FIG. 12, that contains the bases. Those SNP in regions common with v.1 were not shown here. Numbers correspond to those of 98P4B6 v.7. Black box shows the same sequence as 98P4B6 v.7. SNPs are indicated above the box. FIG. 10( c): Schematic alignment of SNP variants of 98P4B6 v.8. Variants 98P4B6 v.25 through v.38 were variants with single nucleotide difference from v.8. Though these SNP variants were shown separately, they could also occur in any combinations and in any transcript variants, as shown in FIG. 12, that contains the bases. Those SNP in regions of common with v.1 were not shown here. Numbers correspond to those of 98P4B6 v.8. Black box shows the same sequence as 98P4B6 v.8. SNPs are indicated above the box.

FIG. 11. Schematic alignment of protein variants of 98P4B6. Protein variants corresponded to nucleotide variants. Nucleotide variants 98P4B6 v.3, v.4, v.9 through v.12, and v.15 through v.19 coded for the same protein as v.1. Nucleotide variants 98P4B6 v.6 and v.8 coded the same protein except for single amino acid at 475, which is an “M” in v.8. Variants v.25 was translated from v.25, a SNP variant of v.8, with one amino acid difference at 565. Similarly, v.21 differed from v.7 by one amino acid at 565. Single amino acid differences were indicated above the boxes. Black boxes represent the same sequence as 98P4B6 v.1. Numbers underneath the box correspond to 98P4B6 v.1.

FIG. 12. Structure of transcript variants of 98P4B6. Variant 98P4B6 v.2 through v.8 were transcript variants of v.1. Variant v.2 was a single exon transcript whose 3′ portion was the same as the last exon of v.1. The first two exons of v.3 were in intron 1 of v. 1. Variants v.4, v.5 and v.6 spliced out 224-334 in the first exon of v.1. In addition, v.5 spliced out exon 5 while v.6 spliced out exon 6 but extended exon 5 of v.1. Variant v.7 used alternative transcription start and different 3′ exons. Variant v.8 extended 5′ end and kept the whole intron 5 of v.1. The first 35 bases of v.1 were not in the nearby 5′ region of v.1 on the current assembly of the human genome. Ends of exons in the transcripts are marked above the boxes. Potential exons of this gene are shown in order as on the human genome. Poly A tails and single nucleotide differences are not shown in the figure. Numbers in “( )” underneath the boxes correspond to those of 98P4B6 v.1. Lengths of introns and exons are not proportional.

FIG. 13. Secondary structure and transmembrane domains prediction for 98P4B6 protein variants. 13(A), 13(B), 13(C), 13(D), 13(E): The secondary structure of 98P4B6 protein variant 1 (SEQ ID NO: 193), Variant 2 (SEQ ID NO: 194), Variant 5 (SEQ ID NO: 195), Variant 6 (SEQ ID NO: 196), and Variant 7 (SEQ ID NO: 197) were predicted using the HNN—Hierarchical Neural Network method, accessed from the ExPasy molecular biology server. This method predicts the presence and location of alpha helices, extended strands, and random coils from the primary protein sequence. The percent of the protein in a given secondary structure is also listed.

13(F), 13(H), 13(J), 13(L), and 13(N): Schematic representations of the probability of existence of transmembrane regions and orientation of 98P4B6 variants 1, 2, 5-7, respectively, based on the TMpred algorithm of Hofmann and Stoffel which utilizes TMBASE (K. Hofmann, W. Stoffel. TMBASE—A database of membrane spanning protein segments Biol. Chem. Hoppe-Seyler 374:166, 1993). 13(G), 13(I), 13(K), 13(M), and 13(O): Schematic representations of the probability of the existence of transmembrane regions and the extracellular and intracellular orientation of 98P486 variants 1, 2, 5-7, respectively, based on the TMHMM algorithm of Sonn hammer, von Heijne, and Krogh (Erik L. L. Sonnhammer, Gunnar von Heijne, and Anders Krogh: A hidden Markov model for predicting transmembrane helices in protein sequences. In Proc. of Sixth Int. Conf. on Intelligent Systems for Molecular Biology, p 175-182 Ed J. Glasgow, T. Littlejohn, F. Major, R. Lathrop, D. Sankoff, and C. Sensen Menlo Park, Calif. AAAI Press, 1998). The TMpred and TMHMM algorithms are accessed from the ExPasy molecular biology server.

FIG. 14. 98P4B6 Expression in Human Normal and Patient Cancer Tissues. First strand cDNA was generated from normal stomach, normal brain, normal heart, normal liver, normal skeletal muscle, normal testis, normal prostate, normal bladder, normal kidney, normal colon, normal lung, normal pancreas, and a pool of cancer specimens from prostate cancer patients; bladder cancer patients, kidney cancer patients, colon cancer patients, lung cancer patients, pancreas cancer patients, and a pool of 2 patient prostate metastasis to lymph node. Normalization was performed by PCR using primers to actin. Semi-quantitative PCR, using primers directed to 98P4B6 v.1, v.13, and v.14 (A), or directed specifically to the splice variants 98P4Bra v.6 and v.8 (B), was performed at 26 and 30 cycles of amplification. Samples were run on an agarose gel, and PCR products were quantitated using the AlphaImager software. Results show strong expression of 98P4B6 v.1, v.13, and v.14 and its splice variants v.6 and v.8 in normal prostate and in prostate cancer. Expression was also detected in bladder cancer, kidney cancer, colon cancer, lung cancer, pancreas cancer, breast cancer, cancer metastasis as well as in the prostate cancer metastasis to lymph node specimens, compared to all normal tissues tested.

FIG. 15. 98P4B6 Expression in lung, ovary, prostate, bladder, cervix, uterus and pancreas patient cancer specimens. First strand cDNA was prepared from a panel of patient cancer specimens. Normalization was performed by PCR using primers to actin. Semi-quantitative PCR, using primers to 98P4B6 v.1, v.13, and v.14, was performed at 26 and 30 cycles of amplification. Samples were run on an agarose gel, and PCR products were quantitated using the AlphaImager software. Expression was recorded as absent, low, medium or strong. Results show expression of 98P4B6 in the majority of all patient cancer specimens tested.

FIG. 16. Expression of 98P4B6 in stomach cancer patient specimens. (A) RNA was extracted from normal stomach (N) and from 10 different stomach cancer patient specimens (T). Northern blot with 10 μg of total RNA/lane was probed with 98P4B6 sequence. Results show strong expression of 98P4B6 in the stomach tumor tissues and lower expression in normal stomach. The lower panel represents ethidium bromide staining of the blot showing quality of the RNA samples. (B) Expression of 98P4B6 was assayed in a panel of human stomach cancers (T) and their respective matched normal tissues (N) on RNA dot blots. 98P4B6 was detected in 7 out of 8 stomach tumors but not in the matched normal tissue.

FIG. 17. Detection of 98P4B6 expression with polyclonal antibody. 293T cells were transfected with 98P4B6.GFP.pcDNA3.1/mychis construct clone A12 or clone B12. STEAP1.GFP vector was used as a positive control. And as a negative control an empty vector was used. Forty hours later, cell lysates were collected. Samples were run on an SDS-PAGE acrylamide gel, blotted and stained with either anti-GFP antibody (A), anti-98P4B6 antibody generated against amino acids 198-389 (B), or anti-98P4B6 antibody generated against amino acids 153-165. The blot was developed using the ECL chemiluminescence kit and visualized by autoradiography. Results show expression of the expected 98P4B6.GFP fusion protein as detected by the anti-GFP antibody. Also, we were able to raise 2 different polyclonal antibodies that recognized the 98P4B6.GFP fusion proteins as shown in B and C.

FIG. 18. Detection of 98P4B6 expression with polyclonal antibody. 293T cells were transfected with 98P4B6.GFP.pcDNA3.1/mychis construct done A12 or done B12. Expression of the 98P4B6.GFP fusion protein was detected by flow cytometry (A) and by fluorescent microscopy (B). Results show strong green fluorescence in the majority of the cells. The fusion protein localized to the perinuclear area and to the cell membrane.

FIG. 19. STEAP-2 Characteristics. The expression of STEAP-2 in normal tissues is predominantly restricted to the prostate. STEAP-2 is expressed in several cancerous tissues. In patient-derived prostate, colon, and lung cancer specimens; and Multiple cancer cell lines, including prostate, colon, Ewing's sarcoma, lung, kidney, pancreas and tests. By ISH, STEAP-2 expression appears to be primarily limited to ductal epithelial cells.

FIG. 20. STEAP-2 Induces Tyrosine Phosphorylation in PC3 Cells. STEAP-2 induces the tyrosine phosphorylation of proteins at 140-150, 120, 75-80, 62 and 40 kDa.

FIG. 21. STEAP-2 Enhances Tyrosine Phosphorylation in NIH 3T3 Cells. STEAP-2 enhances the phosphorylation of p135-140, p78-75 by STEAP-2 in NIH 3T3 cells. STEAP-2 C-Flag enhances the phosphorylation of p180, and induces the de-phosphorylation of p132, p82 and p75.

FIG. 22. STEAP-2 Induces ERK Phosphorylation. STEAP-2 Induces ERK phosphorylation in PC3 and 3T3 cells in 0.5 and 10% FBS. Lack or ERK phosphorylation in 3T3-STEAP-2-cflag cells. Potential role as dominant negative.

FIG. 23. STEAP Enhances Calcium Flux in PC3 cells. PC-STEAP-1 and PC3-STEAP-2 exhibit enhanced calcium flux in response to LPA. PC3-STEAP-1 demonstrates susceptibility to the L type calcium channel inhibitor, conotoxin. PC3-STEAP-2 shown susceptibility to the PQ type calcium channel inhibitor, agatoxin. NDGA and TEA had no effect on the proliferation of PC3-STEAP-2 cells.

FIG. 24. STEAP-2 Alters the Effect of Paclitaxel on PC3 Cells. Other Chemotherapeutics Tested without yielding a differential response between STEAP-expressing and control cells were Flutamide, Genistein, Rapamycin. STEAP-2 confers partial resistance to Paditaxel in PC3 cells. Over 8 fold increase in percent survival of PC3-STEAP-2 relative to PC3-Neo cells.

FIGS. 25A-25C. Inhibition of Apoptosis by STEAP-2. PC3 cells were treated with paclitaxel for 60 hours and analyzed for apoptosis by annexinV-PI staining. Expression of STEAP-2 partially inhibits apoptosis by paclitaxel.

FIGS. 26A-26E. STEAP-2 Attenuates Paclitaxel Mediated Apoptosis. PC3 cells were treated with paclitaxel for 68 hours and analyzed for apoptosis. Expression of STEAP-2, but not STEAP-2CFlag, partially inhibits apoptosis by paclitaxel.

DETAILED DESCRIPTION OF THE INVENTION

Outline of Sections

I.) Definitions

II.) 98P4B6 Polynucleotides

II.A.) Uses of 98P4B6 Polynucleotides

II.A.1.) Monitoring of Genetic Abnormalities

II.A.2.) Antisense Embodiments

II.A.3.) Primers and Primer Pairs

-   -   II.A.4.) Isolation of 98P4B6-Encoding Nucleic Acid Molecules

II.A.5.) Recombinant Nucleic Acid Molecules and Host-Vector Systems

III.) 98P4B6-related Proteins

-   -   III.A.) Motif-bearing Protein Embodiments     -   III.B.) Expression of 98P4B6-related Proteins     -   III.C.) Modifications of 98P4B6-related Proteins     -   III.D.) Uses of 98P4B6-related Proteins

IV.) 98P4B6 Antibodies

V.) 98P4B6 Cellular Immune Responses

VI.) 98P4B6 Transgenic Animals

VII.) Methods for the Detection of 98P4B6

VIII.) Methods for Monitoring the Status of 98P4B6-related Genes and Their Products

IX.) Identification of Molecules That Interact With 98P4B6

X.) Therapeutic Methods and Compositions

-   -   X.A.) Anti-Cancer Vaccines

X.B.) 98P4B6 as a Target for Antibody-Based Therapy

X.C.) 98P4B6 as a Target for Cellular Immune Responses

-   -   X.C.1. Minigene Vaccines     -   X.C.2. Combinations of CTL Peptides with Helper Peptides     -   X.C.3. Combinations of CTL Peptides with T Cell Priming Agents     -   X.C.4. Vaccine Compositions Comprising DC Pulsed with CTL and/or         HTL Peptides     -   X.D.) Adoptive Immunotherapy

X.E.) Administration of Vaccines for Therapeutic or Prophylactic Purposes

XI.) Diagnostic and Prognostic Embodiments of 98P4B6.

XII.) Inhibition of 98P4B6 Protein Function

-   -   XII.A.) Inhibition of 98P4B6 With Intracellular Antibodies     -   XII.B.) Inhibition of 98P4B6 with Recombinant Proteins     -   XII.C.) Inhibition of 98P4B6 Transcription or Translation     -   XII.D.) General Considerations for Therapeutic Strategies

XIII.) Identification, Characterization and Use of Modulators of 98P4B6

XIV.) KITS/Articles of Manufacture

I.) DEFINITIONS

Unless otherwise defined, all terms of art, notations and other scientific terms or terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this invention pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art. Many of the techniques and procedures described or referenced herein are well understood and commonly employed using conventional methodology by those skilled in the art, such as, for example, the widely utilized molecular cloning methodologies described in Sambrook et al., Molecular Cloning: A Laboratory Manual 2nd. edition (1989) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. As appropriate, procedures involving the use of commercially available kits and reagents are generally carried out in accordance with manufacturer defined protocols and/or parameters unless otherwise noted.

The terms “advanced prostate cancer”, “locally advanced prostate cancer”, “advanced disease” and “locally advanced disease” mean prostate cancers that have extended through the prostate capsule, and are meant to include stage C disease under the American Urological Association (AUA) system, stage C1-C2 disease under the Whitmore-Jewett system, and stage T3-T4 and N+ disease under the TNM (tumor, node, metastasis) system. In general, surgery is not recommended for patients with locally advanced disease, and these patients have substantially less favorable outcomes compared to patients having clinically localized (organ-confined) prostate cancer. Locally advanced disease is clinically identified by palpable evidence of induration beyond the lateral border of the prostate, or asymmetry or induration above the prostate base. Locally advanced prostate cancer is presently diagnosed pathologically following radical prostatectomy if the tumor invades or penetrates the prostatic capsule, extends into the surgical margin, or invades the seminal vesicles.

“Altering the native glycosylation pattern” is intended for purposes herein to mean deleting one or more carbohydrate moieties found in native sequence 98P4B6 (either by removing the underlying glycosylation site or by deleting the glycosylation by chemical and/or enzymatic means), and/or adding one or more glycosylation sites that are not present in the native sequence 98P486. In addition, the phrase includes qualitative changes in the glycosylation of the native proteins, involving a change in the nature and proportions of the various carbohydrate moieties present.

The term “analog” refers to a molecule which is structurally similar or shares similar or corresponding attributes with another molecule (e.g. a 98P4B6-related protein). For example, an analog of a 98P4B6 protein can be specifically bound by an antibody or T cell that specifically binds to 98P4B6.

The term “antibody” is used in the broadest sense. Therefore, an “antibody” can be naturally occurring or man-made such as monoclonal antibodies produced by conventional hybridoma technology. Anti-98P4B6 antibodies comprise monoclonal and polyclonal antibodies as well as fragments containing the antigen-binding domain and/or one or more complementarity determining regions of these antibodies.

An “antibody fragment” is defined as at least a portion of the variable region of the immunoglobulin molecule that binds to its target, i.e., the antigen-binding region. In one embodiment it specifically covers single anti-98P4B6 antibodies and clones thereof (including agonist, antagonist and neutralizing antibodies) and anti-98P4B6 antibody compositions with polyepitopic specificity.

The term “codon optimized sequences” refers to nucleotide sequences that have been optimized for a particular host species by replacing any codons having a usage frequency of less than about 20%. Nucleotide sequences that have been optimized for expression in a given host species by elimination of spurious polyadenylation sequences, elimination of exon/intron splicing signals, elimination of transposon-like repeats and/or optimization of GC content in addition to codon optimization are referred to herein as an “expression enhanced sequences.”

A “combinatorial library” is a collection of diverse chemical compounds generated by either chemical synthesis or biological synthesis by combining a number of chemical “building blocks” such as reagents. For example, a linear combinatorial chemical library, such as a polypeptide (e.g., mutein) library, is formed by combining a set of chemical building blocks called amino acids in every possible way for a given compound length (i.e., the number of amino acids in a polypeptide compound). Numerous chemical compounds are synthesized through such combinatorial mixing of chemical building blocks (Gallop et al., J. Med. Chem. 37(9)-1233-1251 (1994)).

Preparation and screening of combinatorial libraries is well known to those of skill in the art. Such combinatorial chemical libraries include, but are not limited to, peptide libraries (see, e.g., U.S. Pat. No. 5,010,175, Furka, Pept. Prot. Res. 37:487-493 (1991), Houghton et al., Nature, 354:84-88 (1991)), peptoids (PCT Publication No WO 91/19735), encoded peptides (PCT Publication WO 93/20242), random bio-oligomers (PCT Publication WO 92/00091), benzodiazepines (U.S. Pat. No. 5,288,514), diversomers such as hydantoins, benzodiazepines and dipeptides (Hobbs et al., Proc. Nat. Acad. Sci. USA 90:6909-6913 (1993)), vinylogous polypeptides (Hagihara et al., J. Amer. Chem. Soc. 114:6568 (1992)), nonpeptidal peptidomimetics with a Beta-D-Glucose scaffolding (Hirschmann et al., J. Amer. Chem. Soc. 114:9217-9218 (1992)), analogous organic syntheses of small compound libraries (Chen et al., J. Amer. Chem. Soc. 116:2661 (1994)), oligocarbamates (Cho, et al., Science 261:1303 (1993)), and/or peptidyl phosphonates (Campbell et al., J. Org. Chem. 59:658 (1994)). See, generally, Gordon et al., J. Med. Chem. 37:1385 (1994), nucleic acid libraries (see, e.g., Stratagene, Corp.), peptide nucleic acid libraries (see, e.g., U.S. Pat. No. 5,539,083), antibody libraries (see, e.g., Vaughn et al., Nature Biotechnology 14(3): 309-314 (1996), and PCT/US96/10287), carbohydrate libraries (see, e.g., Liang et al., Science 274:1520-1522 (1996), and U.S. Pat. No. 5,593,853), and small organic molecule libraries (see, e.g., benzodiazepines, Baum, C&EN, January 18, page 33 (1993); isoprenoids, U.S. Pat. No. 5,569,588; thiazolidinones and metathiazanones, U.S. Pat. No. 5,549,974; pyrrolidines, U.S. Pat. Nos. 5,525,735 and 5,519,134; morpholino compounds, U.S. Pat. No. 5,506,337; benzodiazepines, U.S. Pat. No. 5,288,514; and the like).

Devices for the preparation of combinatorial libraries are commercially available (see, e.g., 357 NIPS, 390 NIPS, Advanced Chem Tech, Louisville Ky.; Symphony, Rainin, Woburn, Mass.; 433A, Applied Biosystems, Foster City, Calif.; 9050, Plus, Millipore, Bedford, NIA). A number of well-known robotic systems have also been developed for solution phase chemistries. These systems include automated workstations such as the automated synthesis apparatus developed by Takeda Chemical Industries, LTD. (Osaka, Japan) and many robotic systems utilizing robotic arms (Zymate H, Zymark Corporation, Hopkinton, Mass.; Orca, Hewlett-Packard, Palo Alto, Calif.), which mimic the manual synthetic operations performed by a chemist. Any of the above devices are suitable for use with the present invention. The nature and implementation of modifications to these devices (if any) so that they can operate as discussed herein will be apparent to persons skilled in the relevant art. In addition, numerous combinatorial libraries are themselves commercially available (see, e.g., ComGenex, Princeton, N.J.; Asinex, Moscow, RU; Tripos, Inc., St. Louis, Mo.; ChemStar, Ltd, Moscow, RU; 3D Pharmaceuticals, Exton, Pa.; Martek Biosciences, Columbia, Md.; etc.).

The term “cytotoxic agent” refers to a substance that inhibits or prevents the expression activity of cells, function of cells and/or causes destruction of cells. The term is intended to include radioactive isotopes chemotherapeutic agents, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof. Examples of cytotoxic agents include, but are not limited to auristatins, auromycins, maytansinoids, yttrium, bismuth, ricin, ricin A-chain, combrestatin, duocarmycins, dolostatins, doxorubicin, daunorubicin, taxol, cisplatin, cc1065, ethidium bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomycin, diphtheria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin, abrin A chain, modeccin A chain, alpha-sarcin, gelonin, mitogellin, restrictocin, phenomycin, enomycin, curicin, crotin, calicheamicin, Sapaonaria officinalis inhibitor, and glucocorticoid and other chemotherapeutic agents, as well as radioisotopes such as At²¹¹, I¹³¹, I¹²⁵, Y⁹⁰, Re¹⁸⁶, Re¹⁸⁸, Sm¹⁵³, Bi^(212 or 213), P³² and radioactive isotopes of Lu including Lu¹⁷⁷. Antibodies may also be conjugated to an anti-cancer pro-drug activating enzyme capable of converting the pro-drug to its active form.

The “gene product” is sometimes referred to herein as a protein or mRNA. For example, a “gene product of the invention” is sometimes referred to herein as a “cancer amino acid sequence”, “cancer protein”, “protein of a cancer listed in Table I”, a “cancer mRNA”, “mRNA of a cancer listed in Table I”, etc. In one embodiment, the cancer protein is encoded by a nucleic acid of FIG. 2. The cancer protein can be a fragment, or alternatively, be the full-length protein to the fragment encoded by the nucleic acids of FIG. 2. In one embodiment, a cancer amino acid sequence is used to determine sequence identity or similarity. In another embodiment, the sequences are naturally occurring allelic variants of a protein encoded by a nucleic acid of FIG. 2. In another embodiment, the sequences are sequence variants as further described herein.

“High throughput screening” assays for the presence, absence, quantification, or other properties of particular nucleic acids or protein products are well known to those of skill in the art. Similarly, binding assays and reporter gene assays are similarly well known. Thus, e.g., U.S. Pat. No. 5,559,410 discloses high throughput screening methods for proteins; U.S. Pat. No. 5,585,639 discloses high throughput screening methods for nucleic acid binding (i.e., in arrays); while U.S. Pat. Nos. 5,576,220 and 5,541,061 disclose high throughput methods of screening for ligand/antibody binding.

In addition, high throughput screening systems are commercially available (see, e.g., Amersham Biosciences, Piscataway, N.J.; Zymark Corp., Hopkinton, Mass.; Air Technical Industries, Mentor, Ohio; Beckman Instruments, Inc. Fullerton, Calif.; Precision Systems, Inc., Natick, Mass.; etc.). These systems typically automate entire procedures, including all sample and reagent pipetting, liquid dispensing, timed incubations, and final readings of the microplate in detector(s) appropriate for the assay. These configurable systems provide high throughput and rapid start up as well as a high degree of flexibility and customization. The manufacturers of such systems provide detailed protocols for various high throughput systems. Thus, e.g., Zymark Corp. provides technical bulletins describing screening systems for detecting the modulation of gene transcription, ligand binding, and the like.

The term “homolog” refers to a molecule which exhibits homology to another molecule, by for example, having sequences of chemical residues that are the same or similar at corresponding positions.

“Human Leukocyte Antigen” or “HLA” is a human class I or class II Major Histocompatibility Complex (MHC) protein (see, e.g., Stites, et al., IMMUNOLOGY, 8^(TH) ED., Lange Publishing, Los Altos, Calif. (1994).

The terms “hybridize”, “hybridizing”, “hybridizes” and the like, used in the context of polynucleotides, are meant to refer to conventional hybridization conditions, preferably such as hybridization in 50% formamide/6×SSC/0.1% SDS/100 μg/ml ssDNA, in which temperatures for hybridization are above 37 degrees C. and temperatures for washing in 0.1×SSC/0.1% SDS are above 55 degrees C.

The phrases “isolated” or “biologically pure” refer to material which is substantially or essentially free from components which normally accompany the material as it is found in its native state. Thus, isolated peptides in accordance with the invention preferably do not contain materials normally associated with the peptides in their in situ environment. For example, a polynucleotide is said to be “isolated” when it is substantially separated from contaminant polynucleotides that correspond or are complementary to genes other than the 98P4B6 genes or that encode polypeptides other than 98P4B6 gene product or fragments thereof. A skilled artisan can readily employ nucleic acid isolation procedures to obtain an isolated 98P4B6 polynucleotide. A protein is said to be “isolated,” for example, when physical, mechanical or chemical methods are employed to remove the 98P4B6 proteins from cellular constituents that are normally associated with the protein. A skilled artisan can readily employ standard purification methods to obtain an isolated 98P4B6 protein. Alternatively, an isolated protein can be prepared by chemical means.

The term “mammal” refers to any organism classified as a mammal, including mice, rats, rabbits, dogs, cats, cows, horses and humans. In one embodiment of the invention, the mammal is a mouse. In another embodiment of the invention, the mammal is a human.

The terms “metastatic prostate cancer” and “metastatic disease” mean prostate cancers that have spread to regional lymph nodes or to distant sites, and are meant to include stage D-disease under the AUA system and stage T×N×M+ under the TNM system. As is the case with locally advanced prostate cancer, surgery is generally not indicated for patients with metastatic disease, and hormonal (androgen ablation) therapy is a preferred treatment modality. Patients with metastatic prostate cancer eventually develop an androgen-refractory state within 12 to 18 months of treatment initiation. Approximately half of these androgen-refractory patients die within 6 months after developing that status. The most common site for prostate cancer metastasis is bone. Prostate cancer bone metastases are often osteoblastic rather than osteolytic (i.e., resulting in net bone formation). Bone metastases are found most frequently in the spine, followed by the femur, pelvis, rib cage, skull and humorous. Other common sites for metastasis include lymph nodes, lung, liver and brain. Metastatic prostate cancer is typically diagnosed by open or laparoscopic pelvic lymphadenectomy, whole body radionuclide scans, skeletal radiography, and/or bone lesion biopsy.

The term “modulator” or “test compound” or “drug candidate” or grammatical equivalents as used herein describe any molecule, e.g., protein, oligopeptide, small organic molecule, polysaccharide, polynucleotide, etc., to be tested for the capacity to directly or indirectly alter the cancer phenotype or the expression of a cancer sequence, e.g., a nucleic acid or protein sequences, or effects of cancer sequences (e.g., signaling, gene expression, protein interaction, etc.) In one aspect, a modulator will neutralize the effect of a cancer protein of the invention. By “neutralize” is meant that an activity of a protein is inhibited or blocked, along with the consequent effect on the cell. In another aspect, a modulator will neutralize the effect of a gene, and its corresponding protein, of the invention by normalizing levels of said protein. In preferred embodiments, modulators alter expression profiles, or expression profile nucleic acids or proteins provided herein, or downstream effector pathways. In one embodiment, the modulator suppresses a cancer phenotype, e.g. to a normal issue fingerprint. In another embodiment, a modulator induced a cancer phenotype. Generally, a plurality of assay mixtures is run in parallel with different agent concentrations to obtain a differential response to the various concentrations. Typically, one of these concentrations serves as a negative control, i.e., at zero concentration or below the level of detection.

Modulators, drug candidates or test compounds encompass numerous chemical classes, though typically they are organic molecules, preferably small organic compounds having a molecular weight of more than 100 and less than about 2,500 Daltons. Preferred small molecules are less than 2000, or less than 1500 or less than 1000 or less than 500 D. Candidate agents comprise functional groups necessary for structural interaction with proteins, particularly hydrogen bonding, and typically include at least an amine, carbonyl, hydroxyl or carboxyl group, preferably at least two of the functional chemical groups. The candidate agents often comprise cyclical carbon or heterocyclic structures and/or aromatic or polyaromatic structures substituted with one or more of the above functional groups. Modulators also comprise biomolecules such as peptides, saccharides, fatty acids, steroids, purines, pyrimidines, derivatives, structural analogs or combinations thereof. Particularly preferred are peptides. One class of modulators are peptides, for example of from about five to about 35 amino acids, with from about five to about 20 amino acids being preferred, and from about 7 to about 15 being particularly preferred. Preferably, the cancer modulatory protein is soluble, includes a don-transmembrane region, and/or, has an N-terminal Cys to aid in solubility. In one embodiment, the C-terminus of the fragment is kept as a free add and the N-terminus is a free amine to aid in coupling, i.e., to cysteine. In one embodiment, a cancer protein of the invention is conjugated to an immunogenic agent as discussed herein. In one embodiment, the cancer protein is conjugated to BSA. The peptides of the invention, e.g., of preferred lengths, can be linked to each other or to other amino adds to create a longer peptide/protein. The modulatory peptides can be digests of naturally occurring proteins as is outlined above, random peptides, or “biased” random peptides. In a preferred embodiment, peptide/protein-based modulators are antibodies, and fragments thereof, as defined herein.

Modulators of cancer can also be nucleic acids. Nucleic acid modulating agents can be naturally occurring nucleic acids, random nucleic adds, or “biased” random nucleic acids. For example, digests of prokaryotic or eukaryotic genomes can be used in an approach analogous to that outlined above for proteins.

The term “monoclonal antibody” refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the antibodies comprising the population are identical except for possible naturally occurring mutations that are present in minor amounts.

A “motif”, as in biological motif of a 98P4B6-related protein, refers to any pattern of amino adds forming part of the primary sequence of a protein, that is associated with a particular function (e.g. protein-protein interaction, protein-DNA interaction, etc) or modification (e.g. that is phosphorylated, glycosylated or amidated), or localization (e.g. secretory sequence, nuclear localization sequence, etc.) or a sequence that is correlated with being immunogenic, either humorally or cellularly. A motif can be either contiguous or capable of being aligned to certain positions that are generally correlated with a certain function or property. In the context of HLA motifs, “motif” refers to the pattern of residues in a peptide of defined length, usually a peptide of from about 8 to about 13 amino adds for a class I HLA motif and from about 6 to about 25 amino acids for a class II HLA motif, which is recognized by a particular HLA molecule. Peptide motifs for HLA binding are typically different for each protein encoded by each human HLA allele and differ in the pattern of the primary and secondary anchor residues.

A “pharmaceutical excipient” comprises a material such as an adjuvant, a carrier, pH-adjusting and buffering agents, tonicity adjusting agents, wetting agents, preservative, and the like.

“Pharmaceutically acceptable” refers to a non-toxic, inert, and/or composition that is physiologically compatible with humans or other mammals.

The term “polynucleotide” means a polymeric form of nucleotides of at least 10 bases or base pairs in length, either ribonucleotides or deoxynucleotides or a modified form of either type of nucleotide, and is meant to include single and double stranded forms of DNA and/or RNA. In the art, this term if often used interchangeably with “oligonucleotide”. A polynucleotide can comprise a nucleotide sequence disclosed herein wherein thymidine (T), as shown for example in FIG. 2, can also be uracil (U); this definition pertains to the differences between the chemical structures of DNA and RNA, in particular the observation that one of the four major bases in RNA is uracil (U) instead of thymidine (T).

The term “polypeptide” means a polymer of at least about 4, 5, 6, 7, or 8 amino acids. Throughout the specification, standard three letter or single letter designations for amino acids are used. In the art, this term is often used interchangeably with “peptide” or “protein”.

An HLA “primary anchor residue” is an amino acid at a specific position along a peptide sequence which is understood to provide a contact point between the immunogenic peptide and the HLA molecule. One to three, usually two, primary anchor residues within a peptide of defined length generally defines a “motif” for an immunogenic peptide. These residues are understood to fit in close contact with peptide binding groove of an HLA molecule, with their side chains buried in specific pockets of the binding groove. In one embodiment, for example, the primary anchor residues for an HLA class I molecule are located at position 2 (from the amino terminal position) and at the carboxyl terminal position of a 8, 9, 10, 11, or 12 residue peptide epitope in accordance with the invention. Alternatively, in another embodiment, the primary anchor residues of a peptide binds an HLA class II molecule are spaced relative to each other, rather than to the termini of a peptide, where the peptide is generally of at least 9 amino acids in length. The primary anchor positions for each motif and supermotif are set forth in Table IV. For example, analog peptides can be created by altering the presence or absence of particular residues in the primary and/or secondary anchor positions shown in Table IV. Such analogs are used to modulate the binding affinity and/or population coverage of a peptide comprising a particular HLA motif or supermotif.

“Radioisotopes” include, but are not limited to the following (non-limiting exemplary uses are also set forth):

Examples of Medical Isotopes

Isotope

Description of use

Actinium-225

(AC-225)

See Thorium-229 (Th-229)

Actinium-227

(AC-227)

Parent of Radium-223 (Ra-223) which is an Alpha Emitter Used to Treat Metastases in the Skeleton Resulting from cancer (i.e., breast and prostate cancers), and cancer radioimmunotherapy

Bismuth-212

(Bi-212)

See Thorium-228 (Th-228)

Bismuth-213

(Bi-213)

See Thorium-229 (Th-229)

Cadmium-109

(Cd-109)

Cancer detection

Cobalt-60

(Co-60)

Radiation source for radiotherapy of cancer, for food irradiators, and for sterilization of medical supplies

Copper-64

(Cu-64)

A positron emitter used for cancer therapy and SPECT imaging

Copper-67

(Cu-67)

Beta/gamma emitter used in cancer radioimmunotherapy and diagnostic studies (i.e., breast and colon cancers, and lymphoma)

Dysprosium-166

(Dy-166)

Cancer radioimmunotherapy

Erbium-169

(Er-169)

Rheumatoid arthritis treatment, particularly for the small joints associated with fingers and toes

Europium-152

(Eu-152)

Radiation source for food irradiation and for sterilization of medical supplies Europium-154

(Eu-154)

Radiation source for food irradiation and for sterilization of medical supplies

Gadolinium-153

(Gd-153)

Osteoporosis detection and nuclear medical quality assurance devices

Gold-198

(Au-198)

Implant and intracavity therapy of ovarian, prostate, and brain cancers

Holmium-166

(Ho-166)

Multiple myeloma treatment in targeted skeletal therapy, cancer radioimmunotherapy, bone marrow ablation, and rheumatoid arthritis treatment

Iodine-125

(I-125)

Osteoporosis detection, diagnostic imaging, tracer drugs, brain cancer treatment, radiolabeling, tumor imaging, mapping of receptors in the brain, interstitial radiation therapy, brachytherapy for treatment of prostate cancer, determination of glomerular filtration rate (GFR), determination of plasma volume, detection of deep vein thrombosis of the legs Iodine-131 (I-131) Thyroid function evaluation, thyroid disease detection, treatment of thyroid cancer as well as other non-malignant thyroid diseases (i.e., Graves disease, goiters, and hyperthyroidism), treatment of leukemia, lymphoma, and other forms of cancer (e.g., breast cancer) using radioimmunotherapy Iridium-192 (Ir-192) Brachytherapy, brain and spinal cord tumor treatment, treatment of blocked arteries (i.e., arteriosclerosis and restenosis), and implants for breast and prostate tumors Lutetium-177 (Lu-177) Cancer radioimmunotherapy and treatment of blocked arteries (i.e., arteriosclerosis and restenosis) Molybdenum-99 (Mo-99) Parent of Technetium-99m (Tc-99m) which is used for imaging the brain, liver, lungs, heart, and other organs. Currently, Tc-99m is the most widely used radioisotope used for diagnostic imaging of various cancers and diseases involving the brain, heart, liver, lungs; also used in detection of deep vein thrombosis of the legs Osmium-194 (Os-194) Cancer radioimmunotherapy Palladium-103 (Pd-10 3). Prostate cancer treatment Platinum-195m (Pt-195m) Studies on biodistribution and metabolism of cisplatin, a chemotherapeutic drug Phosphorus-32 (P-32) Polycythemia Rubra Vera (Blood Cell Disease) and Leukemia Treatment, Bone Cancer Diagnosis/Treatment; Colon, pancreatic, and liver cancer treatment; radiolabeling nucleic acids for in vitro research, diagnosis of superficial tumors, treatment of blocked arteries (i.e., arteriosclerosis and restenosis), and intracavity therapy Phosphorus-33 (P-33) Leukemia Treatment, Bone Disease Diagnosis/Treatment, Radiolabeling, and Treatment of Blocked Arteries (I.E., arteriosclerosis and restenosis) Radium-223 (Ra-223) See Actinium-227 (Ac-227) Rhenium-186 (Re-186) Bone Cancer Pain Relief, Rheumatoid Arthritis Treatment, and Diagnosis and Treatment of Lymphoma and Bone, breast, colon, and liver cancers using radioimmunotherapy Rhenium-188 (Re-188) Cancer Diagnosis and Treatment Using Radioimmunotherapy, Bone Cancer Pain Relief, Treatment of Rheumatoid arthritis, and treatment of prostate cancer Rhodium-105 (Rh-105) Cancer Radioimmunotherapy Samarium-145 (Sm-145) Ocular cancer treatment Samarium-153 (Sm-153) Cancer radioimmunotherapy and bone cancer pain relief Scandium-47 (Sc-47) Cancer radioimmunotherapy and bone cancer pain relief Selenium-75 (Se-75) Radiotracer used in brain studies, imaging of adrenal cortex by gamma-scintigraphy, lateral locations of steroid secreting tumors, pancreatic scanning, detection of hyperactive parathyroid glands, measure rate of bile acid loss from the endogenous pool Strontum-85 (Sr-85) Bone cancer detection and brain scans Strontium-89 (Sr-89) Bone cancer pain relief, multiple myeloma treatment, and osteoblastic therapy Technetium-99m (Tc-99M) See Molybdenum-99 (Mo-99) Thorium-228 (Th-228) Parent of Bismuth-212 (Bi-212) which is an alpha emitter used in cancer radioimmunotherapy Thorium-229 (Th-229) Parent of Actinium-225 (Ac-225) and grandparent of Bismuth-213 (Bi-213) which are alpha emitters used in cancer radioimmunotherapy Thulium-170 (Tm-170) Gamma source for blood irradiators, energy source for implanted medical devices Tin-117m (Sn-117m) Cancer immunotherapy and bone cancer pain relief Tungsten-188 (W-188) Parent for Rhenium-188 (Re-188) which is Used for Cancer Diagnostics/Treatment, Bone Cancer Pain Relief, rheumatoid arthritis treatment, and treatment of blocked arteries (i.e., arteriosclerosis and restenosis) Xenon-127 (Xe-127) Neuroimaging of brain disorders, high resolution SPECT studies, pulmonary function tests, and cerebral blood flow studies Ytterbium-175 (Yb-175) Cancer radioimmunotherapy Yttnium-90 (Y-90) Microseeds Obtained from Irradiating Yttrium-89 (Y-89) for Liver Cancer Treatment Yttrium-91 (Y-91) A gamma-emitting label for Yttrium-90 (Y-90) which is used for cancer radioimmunotherapy. (i.e., lymphoma, breast, colon, kidney, lung, ovarian, prostate, pancreatic, and inoperable liver cancers)

By “randomized” or grammatical equivalents as herein applied to nucleic acids and proteins is meant that each nucleic acid and peptide consists of essentially random nucleotides and amino acids, respectively. These random peptides (or nucleic acids, discussed herein) can incorporate any nucleotide or amino acid at any position. The synthetic process can be designed to generate randomized proteins or nucleic acids, to allow the formation of all or most of the possible combinations over the length of the sequence, thus forming a library of randomized candidate bioactive proteinaceous agents.

In one embodiment a library is “fully randomized,” with no sequence preferences or constants at any position. In another embodiment, the library is a “biased random” library. That is, some positions within the sequence either are held constant, or are selected from a limited number of possibilities. For example, the nucleotides or amino acid residues are randomized within a defined class, e.g., of hydrophobic amino adds, hydrophilic residues, sterically biased (either small or large) residues, towards the creation of nucleic acid binding domains, the creation of cysteines, for cross-linking, prolines for SH-3 domains, serines, threonines, tyrosines or histidines for phosphorylation sites, etc., or to purines, etc.

A “recombinant” DNA or RNA molecule is a DNA or RNA molecule that has been subjected to molecular manipulation in vitro.

Non-limiting examples of small molecules include compounds that bind or interact with 98P4B6, ligands including hormones, neuropeptides, chemokines, odorants, phospholipids, and functional equivalents thereof that bind and preferably inhibit 98P4B6 protein function. Such non-limiting small molecules preferably have a molecular weight of less than about 10 kDa, more preferably below about 9, about 8, about 7, about 6, about 5 or about 4 kDa. In certain embodiments, small molecules physically associate with, or bind, 98P4B6 protein; are not found in naturally occurring metabolic pathways; and/or are more soluble in aqueous than non-aqueous solutions

“Stringency” of hybridization reactions is readily determinable by one of ordinary skill in the art, and generally is an empirical calculation dependent upon probe length, washing temperature, and salt concentration. In general, longer probes require higher temperatures for proper annealing, while shorter probes need lower temperatures. Hybridization generally depends on the ability of denatured nucleic acid sequences to reanneal when complementary strands are present in an environment below their melting temperature. The higher the degree of desired homology between the probe and hybridizable sequence, the higher the relative temperature that can be used. As a result, it follows that higher relative temperatures would tend to make the reaction conditions more stringent, while lower temperatures less so. For additional details and explanation of stringency of hybridization reactions, see Ausubel et al., Current Protocols in Molecular Biology, Wiley Interscience Publishers, (1995).

“Stringent conditions” or “high stringency conditions”, as defined herein, are identified by, but not limited to, those that: (1) employ low ionic strength and high temperature for washing, for example 0.015 M sodium chloride/0.0015 M sodium citrate/0.1% sodium dodecyl sulfate at 50° C.; (2) employ during hybridization a denaturing agent, such as formamide, for example, 50% (v/v) formamide with 0.1% bovine serum albumin/0.1% Ficoll/0.1% polyvinylpyrrolidone/50 mM sodium phosphate buffer at pH 6.5 with 750 mM sodium chloride, 75 mM sodium citrate at 42° C.; or (3) employ 50% formamide, 5×SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5×Denhardt's solution, sonicated salmon sperm DNA (50 μg/ml), 0.1% SDS, and 10% dextran sulfate at 42° C., with washes at 42° C. in 0.2×SSC (sodium chloride/sodium citrate) and 50% formamide at 55° C., followed by a high-stringency wash consisting of 0.1×SSC containing EDTA at 55° C. “Moderately stringent conditions” are described by, but not limited to, those in Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Press, 1989, and include the use of washing solution and hybridization conditions (e.g., temperature, ionic strength and % SDS) less stringent than those described above. An example of moderately stringent conditions is overnight incubation at 37° C. in a solution comprising: 20% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 mg/mL denatured sheared salmon sperm DNA, followed by washing the filters in 1×SSC at about 37-50° C. The skilled artisan will recognize how to adjust the temperature, ionic strength, etc. as necessary to accommodate factors such as probe length and the like.

An HLA “supermotif” is a peptide binding specificity shared by HLA molecules encoded by two or more HLA alleles. Overall phenotypic frequencies of HLA-supertypes in different ethnic populations are set forth in Table IV (F). The non-limiting constituents of various supertypes are as follows:

A2: A*0201, A*0202, A*0203, A*0204, A*0205, A*0206, A*6802, A*6901, A*0207

A3: A3, A11, A31, A*3301, A*6801, A*0301, A*1101, A*3101

B7: B7, B*3501-03, B*51, B*5301, B*5401, B*5501, B*5502, B*5601, B*6701, B*7801, B*0702, B*5101, B*5602

B44: B*3701, B*4402, B*4403, B*60 (B*4001), B61 (B*4006)

A1: A*0102, A*2604, A*3601, A*4301, A*8001

A24: A*24, A*30, A*2403, A*2404, A*3002, A*3003

B27: B*1401-02, B*1503, B*1509, B*1510, B*1518, B*3801-02, B*3901, B*3902, B*3903-04, B*4801-02, B*7301, B*2701-08

B58: B*1516, B*1517, B*5701, B*5702, B58

B62: B*4601. B52, B*1501 (B62), B*1502 (B75), B*1513 (B77)

Calculated population coverage afforded by different HLA-supertype combinations are set forth in Table IV (G).

As used herein “to treat” or “therapeutic” and grammatically related terms, refer to any improvement of any consequence of disease, such as prolonged survival, less morbidity, and/or a lessening of side effects which are the byproducts of an alternative therapeutic modality; full eradication of disease is not required.

A “transgenic animal” (e.g., a mouse or rat) is an animal having cells that contain a transgene, which transgene was introduced into the animal or an ancestor of the animal at a prenatal, e.g., an embryonic stage. A “transgene” is a DNA that is integrated into the genome of a cell from which a transgenic animal develops.

As used herein, an HLA or cellular immune response “vaccine” is a composition that contains or encodes one or more peptides of the invention. There are numerous embodiments of such vaccines, such as a cocktail of one or more individual peptides; one or more peptides of the invention comprised by a polyepitopic peptide; or nucleic acids that encode such individual peptides or polypeptides, e.g., a minigene that encodes a polyepitopic peptide. The “one or more peptides” can include any whole unit integer from 1-150 or more, e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, or 150 or more peptides of the invention. The peptides or polypeptides can optionally be modified, such as by lipidation, addition of targeting or other sequences. HLA class I peptides of the invention can be admixed with, or linked to, HLA class II peptides, to facilitate activation of both cytotoxic T lymphocytes and helper T lymphocytes. HLA vaccines can also comprise peptide-pulsed antigen presenting cells, e.g., dendritic cells.

The term “variant” refers to a molecule that exhibits a variation from a described type or norm, such as a protein that has one or more different amino acid residues in the corresponding position(s) of a specifically described protein (e.g. the 98P4B6 protein shown in FIG. 2 or FIG. 3. An analog is an example of a variant protein. Splice isoforms and single nucleotides polymorphisms (SNPs) are further examples of variants.

The “98P4B6-related proteins” of the invention include those specifically identified herein, as well as allelic variants, conservative substitution variants, analogs and homologs that can be isolated/generated and characterized without undue experimentation following the methods outlined herein or readily available in the art. Fusion proteins that combine parts of different 98P486 proteins or fragments thereof, as well as fusion proteins of a 98P4B6 protein and a heterologous polypeptide are also included. Such 98P4B6 proteins are collectively referred to as the 98P4B6-related proteins, the proteins of the invention, or 98P4B6. The term “98P4B6-related protein” refers to a polypeptide fragment or a 98P4B6 protein sequence of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more than 25 amino adds; or, at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450; 475, 500, 525, 550, 575, or 576 or more amino acids.

II.) 98P4B6 POLYNUCLEOTIDES

One aspect of the invention provides polynucleotides corresponding or complementary to all or part of a 98P4B6 gene, mRNA, and/or coding sequence, preferably in isolated form, including polynucleotides encoding a 98P4B6-related protein and fragments thereof, DNA, RNA, DNA/RNA hybrid, and related molecules, polynucleotides or oligonucleotides complementary to a 98P4B6 gene or mRNA sequence or a part thereof, and polynucleotides or oligonucleotides that hybridize to a 98P4B6 gene, mRNA, or to a 98P4B6 encoding polynucleotide (collectively, “98P486 polynucleotides”). In all instances when referred to in this section, T can also be U in FIG. 2.

Embodiments of a 98P4B6 polynucleotide include: a 98P4B6 polynucleotide having the sequence shown in FIG. 2, the nucleotide sequence of 98P4B6 as shown in FIG. 2 wherein T is U; at least 10 contiguous nucleotides of a polynucleotide having the sequence as shown in FIG. 2; or, at least 10 contiguous nucleotides of a polynucleotide having the sequence as shown in FIG. 2 where T is U. For example, embodiments of 98P4B6 nucleotides comprise, without limitation:

-   -   (I) a polynucleotide comprising, consisting essentially of, or         consisting of a sequence as shown in FIG. 2, wherein T can also         be U;     -   (II) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2A, from nucleotide         residue number 355 through nucleotide residue number 1719,         including the stop codon, wherein T can also be U;     -   (III) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2B, from nudeotide         residue number 4 through nucleotide residue number 138,         including the stop codon, wherein T can also be U;     -   (IV) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2C, from nucleotide         residue number 188 through nucleotide residue number 1552,         including the a stop codon, wherein T can also be U;     -   (V) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2D, from nucleotide         residue number 318 through nucleotide residue number 1682,         including the stop codon, wherein T can also be U;     -   (VI) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2E, from nucleotide         residue number 318 through nucleotide residue number 1577,         including the stop codon, wherein T can also be U;     -   (VII) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2F, from nucleotide         residue number 318 through nucleotide residue number 1790,         including the stop codon, wherein T can also be U;     -   (VIII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2G, from         nucleotide residue number 295 through nucleotide residue number         2025, including the stop codon, wherein T can also be U;     -   (IX) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2H, from nucleotide         residue number 394 through nucleotide residue number 1866,         including the stop codon, wherein T can also be U;     -   (X) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2 i, from nucleotide         residue number 355 through nucleotide residue number 1719,         including the stop codon, wherein T can also be U;     -   (XI) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2J, from nucleotide         residue number 355 through nucleotide residue number 1719,         including the stop codon, wherein T can also be U;     -   (XII) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2K, from nucleotide         residue number 355 through nucleotide residue number 1719,         including the stop codon, wherein T can also be U;     -   (XIII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2L, from         nucleotide residue number 355 through nucleotide residue number         1719, including the stop codon, wherein T can also be U;     -   (XIV) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2M, from nucleotide         residue number 355 through nucleotide residue number 1719,         including the stop codon, wherein T can also be U;     -   (XV) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2N, from nucleotide         residue number 355 through nucleotide residue number 1719,         including the stop codon, wherein T can also be U;     -   (XVI) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2O, from nucleotide         residue number 355 through nucleotide residue number 1719,         including the stop codon, wherein T can also be U;     -   (XVII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2P, from         nucleotide residue number 355 through nucleotide residue number         1719, including the stop codon, wherein T can also be U;     -   (XVIII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2Q, from         nucleotide residue number 355 through nucleotide residue number         1719, including the stop codon, wherein T can also be U;     -   (XIX) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2R, from nucleotide         residue number 355 through nucleotide residue number 1719,         including the stop codon, wherein T can also be U;     -   (XX) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2S, from nucleotide         residue number 355 through nucleotide residue number 1719,         including the stop codon, wherein T can also be U;     -   (XXI) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2T, from nucleotide         residue number 295 through nucleotide residue number 2025,         including the stop codon, wherein T can also be U;     -   (XXII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2U, from         nucleotide residue number 295 through nucleotide residue number         2025, including the stop codon, wherein T can also be U;     -   (XXIII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2V, from         nucleotide residue number 295 through nucleotide residue number         2025, including the stop codon, wherein T can also be U;     -   (XXIV) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2W, from         nucleotide residue number 295 through nucleotide residue number         2025, including the stop codon, wherein T can also be U;     -   (XXV) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2X, from nucleotide         residue number 295 through nucleotide residue number 2025,         including the stop codon, wherein T can also be U;     -   (XXVI) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2Y, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXVII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2Z, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXVIII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2AA, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXIX) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2AB, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXX) a polynucleotide comprising, consisting essentially of, or         consisting of the sequence as shown in FIG. 2AC, from nucleotide         residue number 394 through nucleotide residue number 1866,         including the stop codon, wherein T can also be U;     -   (XXXI) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2AD, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXXII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2AE, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXXIII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2AF, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXIV) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2AG, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXXV) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2AH, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXXVI) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2AI, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   XXXVII) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2AJ, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXXVIII) a polynucleotide comprising, consisting essentially         of, or consisting of the sequence as shown in FIG. 2AK, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XXXIX) a polynucleotide comprising, consisting essentially of,         or consisting of the sequence as shown in FIG. 2AL, from         nucleotide residue number 394 through nucleotide residue number         1866, including the stop codon, wherein T can also be U;     -   (XL) a polynucleotide that encodes a 98P4B6-related protein that         is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%         homologous to an entire amino acid sequence shown in FIG. 2A-AL;     -   (XLI) a polynucleotide that encodes a 98P4B6-related protein         that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%         identical to an entire amino acid sequence shown in FIG. 2A-AL;     -   (XLII) a polynucleoide that encodes at least one peptide set         forth in Tables VIII-XXI and XXII-XLIX;     -   (XLIII) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIGS. 3A, 3G, and 3H in any whole number         increment up to 454 that includes at least 1, 2, 3, 4, 5, 6, 7,         8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,         24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid         position(s) having a value greater than 0.5 in the         Hydrophilicity profile of FIG. 5;     -   (XLIV) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIGS. 3A, 3G, and 3H in any whole number         increment up to 454 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value less than 0.5 in the Hydropathicity profile of FIG. 6;     -   (XLV) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIGS. 3A, 3G, and 3H in any whole number         increment up to 454 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value greater than 0.5 in the Percent Accessible Residues         profile of FIG. 7;     -   (XLVI) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         adds of a peptide of FIGS. 3A, 3G, and 3H in any whole number         increment up to 454 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value greater than 0.5 in the Average Flexibility profile of         FIG. 8;     -   (XLVII) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIGS. 3A, 3G, and 3H in any whole number         increment up to 454 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value greater than 0.5 in the Beta-turn profile of FIG. 9;     -   (XLVIII) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIG. 3B in any whole number increment up         to 45 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,         14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,         30, 31, 32, 33, 34, 35 amino acid position(s) having a value         greater than 0.5 in the Hydrophilicity profile of FIG. 5;     -   (XLIX) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIG. 3B in any whole number increment up         to 45 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,         14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,         30, 31, 32, 33, 34, 35 amino acid position(s) having a value         less than 0.5 in the Hydropathicity profile of FIG. 6;     -   (L) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIG. 3B in any whole number increment up         to 45 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,         14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,         30, 31, 32, 33, 34, 35 amino acid position(s) having a value         greater than 0.5 in the Percent Accessible Residues profile of         FIG. 7;     -   (LI) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIG. 3B in any whole number increment up         to 45 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,         14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,         30, 31, 32, 33, 34, 35 amino acid position(s) having a value         greater than 0.5 in the Average Flexibility profile of FIG. 8;     -   (LII) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIG. 3B in any whole number increment up         to 45 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,         14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,         30, 31, 32, 33, 34, 35 amino acid position(s) having a value         greater than 0.5 in the Beta-turn profile of FIG. 9     -   (LIII) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIG. 3C in any whole number increment up         to 419 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,         14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,         30, 31, 32, 33, 34, 35 amino acid position(s) having a value         greater than 0.5 in the Hydrophilicity profile of FIG. 5;     -   (LIV) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIG. 3C in any whole number increment up         to 419 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,         14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,         30, 31, 32, 33, 34, 35 amino acid position(s) having a value         less than 0.5 in the Hydropathicity profile of FIG. 6;     -   (LV) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIG. 3C in any whole number increment up         to 419 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,         14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,         30, 31, 32, 33; 34, 35 amino acid position(s) having a value         greater than 0.5 in the Percent Accessible Residues profile of         FIG. 7;     -   (LVI) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIG. 3C in any whole number increment up         to 419 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,         14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,         30, 31, 32, 33, 34, 35 amino acid position(s) having a value         greater than 0.5 in the Average Flexibility profile of FIG. 8;     -   (LVII) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIG. 3C in any whole number increment up         to 419 that includes 1; 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,         14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,         30, 31, 32, 33, 34, 35 amino acid position(s) having a value         greater than 0.5 in the Beta-turn profile of FIG. 9     -   (LVIII) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         adds of a peptide of FIGS. 3D, 3F, and 3J in any whole number         increment up to 490 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value greater than 0.5 in the Hydrophilicity profile of FIG.         5;     -   (LIX) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         adds of a peptide of FIGS. 3D, 3F, and 3J in any whole number         increment up to 490 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value less than 0.5 in the Hydropathicity profile of FIG. 6;     -   (LX) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIGS. 3D, 3F, and 3J in any whole number         increment up to 490 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value greater than 0.5 in the Percent Accessible Residues         profile of FIG. 7;     -   (LXI) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         adds of a peptide of FIGS. 3D, 3F, and 3J in any whole number         increment up to 490 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino'acid position(s) having         a value greater than 0.5 in the Average Flexibility profile of         FIG. 8;     -   (LXII) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIGS. 3D, 3F, and 3J in any whole number         increment up to 490 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value greater than 0.5 in the Beta-turn profile of FIG. 9     -   (LXIII) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIGS. 3E and 3I in any whole number         increment up to 576 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value greater than 0.5 in the Hydrophilicity profile of FIG.         5;     -   (LXIV) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         adds of a peptide of FIGS. 3E and 3I in any whole number         increment up to 576 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value less than 0.5 in the Hydropathicity profile of FIG. 6;     -   (LXV) a polynucleotide that encodes a peptide region of at least         5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,         22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         adds of a peptide of FIGS. 3E and 3I in any whole number         increment up to 576 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value greater than 0.5 in the Percent Accessible Residues         profile of FIG. 7;     -   (LXVI) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIGS. 3E and 3I in any whole number         increment up to 576 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value greater than 0.5 in the Average Flexibility profile of         FIG. 8;     -   (LXVII) a polynucleotide that encodes a peptide region of at         least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,         21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino         acids of a peptide of FIGS. 3E and 3I in any whole number         increment up to 576 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,         11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,         27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having         a value greater than 0.5 in the Beta-turn profile of FIG. 9     -   (LXVIII) a polynucleotide that is fully complementary to a         polynucleotide of any one of (I)-(LXVII).     -   (LXIX) a peptide that is encoded by any of (I) to (LXVIII); and     -   (LXX) a composition comprising a polynucleotide of any of         (1)-(LXVIII) or peptide of (LXIX) together with a pharmaceutical         excipient and/or in a human unit dose form.     -   (LXXI) a method of using a polynucleotide of any (I)-(LXVIII) or         peptide of (LXIX) or a composition of (LXX) in a method to         modulate a cell expressing 98P4B6,     -   (LXXII) a method of using a polynucleotide of any (I)-(LXVIII)         or peptide of (LXIX) or a composition of (LXX) in a method to         diagnose, prophylax, prognose, or treat an individual who bears         a cell expressing 98P4B6     -   (LXXIII) a method of using a polynucleotide of any (I)-(LXVIII)         or peptide of (LXIX) or a composition of (LXX) in a method to         diagnose, prophylax, prognose, or treat an individual who bears         a cell expressing 98P4B6, said cell from a cancer of a tissue         listed in Table I;     -   (LXXIV) a method of using a polynucleotide of any (I)-(LXVIII)         or peptide of (LXIX) or a composition of (LXX) in a method to         diagnose, prophylax, prognose, or treat a a cancer;     -   (LXXV) a method of using a polynucleotide of any (I)-(LXVIII) or         peptide of (LXIX) or a composition of (LXX) in a method to         diagnose, prophylax, prognose, or treat a a cancer of a tissue         listed in Table I; and,     -   (LXXVI) a method of using a polynucleotide of any (I)-(LXVIII)         or peptide of (LXIX) or a composition of (LXX) in a method to         identify or characterize a modulator of a cell expressing         98P4B6.

As used herein, a range is understood to disclose specifically all whole unit positions thereof.

Typical embodiments of the invention disclosed herein include 98P4B6 polynucleotides that encode specific portions of 98P4B6 mRNA sequences-(and those which are complementary to such sequences) such as those that encode the proteins and/or fragments thereof, for example:

(a) 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 225, 250, 275, 300, 325, 350, 375, 400, 410, 420, 430, 440, 450 or 454 or more contiguous amino adds of 98P4B6 variant 1; the maximal lengths relevant for other variants are: variant 2, 44 amino adds; variant 5, 419 amino adds, variant 6, 490 amino adds, variant 7, 576 amino adds, variant 8, 490 amino adds, variant 13, 454 amino adds; variant 14, 454 amino adds, variant 21, 576 amino adds, and variant 25, 490 amino adds.

For example, representative embodiments of the invention disclosed herein include: polynucleotides and their encoded peptides themselves encoding about amino acid 1 to about amino acid 10 of the 98P4B6 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 10 to about amino acid 20 of the 98P4B6 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 20 to about amino acid 30 of the 98P4B6 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 30 to about amino acid 40 of the 98P4B6 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 40 to about amino acid 50 of the 98P4B6 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 50 to about amino acid 60 of the 98P4B6 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 60 to about amino acid 70 of the 98P4B6 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 70 to about amino acid 80 of the 98P4B6 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 80 to about amino acid 90 of the 98P4B6 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 90 to about amino acid 100 of the 98P4B6 protein shown in FIG. 2 or FIG. 3, in increments of about 10 amino adds, ending at the carboxyl terminal amino acid set forth in FIG. 2 or FIG. 3. Accordingly, polynucleotides encoding portions of the amino acid sequence (of about 10 amino acids), of amino acids, 100 through the carboxyl terminal amino acid of the 98P4B6 protein are embodiments of the invention. Wherein it is understood that each particular amino acid position discloses that position plus or minus five amino acid residues.

Polynucleotides encoding relatively long portions of a 98P4B6 protein are also within the scope of the invention. For example, polynucleotides encoding from about amino acid 1 (or 20 or 30 or 40 etc.) to about amino acid 20, (or 30, or 40 or 50 etc.) of the 98P4B6 protein “or variant” shown in FIG. 2 or FIG. 3 can be generated by a variety of techniques well known in the art. These polynucleotide fragments can include any portion of the 98P4B6 sequence as shown in FIG. 2.

Additional illustrative embodiments of the invention disclosed herein include 98P4B6 polynucleotide fragments encoding one or more of the biological motifs contained within a 98P4B6 protein “or variant” sequence, including one or more of the motif-bearing subsequences of a 98P4B6 protein “or variant” set forth in Tables VIII-XXI and XXII-XLIX. In another embodiment, typical polynucleotide fragments of the invention encode one or more of the regions of 98P4B6 protein or variant that exhibit homology to a known molecule. In another embodiment of the invention, typical polynucleotide fragments can encode one or more of the 98P4B6 protein or variant N-glycosylation sites, cAMP and cGMP-dependent protein kinase phosphorylation sites, casein kinase II phosphorylation sites or N-myristoylation site and amidation sites.

Note that to determine the starting position of any peptide set forth in Tables VIII-XXI and Tables XXII to XLIX (collectively HLA Peptide Tables) respective to its parental protein, e.g., variant 1, variant 2, etc., reference is made to three factors: the particular variant, the length of the peptide in an HLA Peptide Table, and the Search Peptides listed in Table VII. Generally, a unique Search Peptide is used to obtain HLA peptides for a particular variant. The position of each Search Peptide relative to its respective parent molecule is listed in Table VII. Accordingly, if a Search Peptide begins at position “X”, one must add the value “X minus 1” to each position in Tables VIII-XXI and Tables XXII-IL to obtain the actual position of the HLA peptides in their parental molecule. For example if a particular Search Peptide begins at position 150 of its parental molecule, one must add 150−1, i.e., 149 to each HLA peptide amino acid position to calculate the position of that amino acid in the parent molecule.

II.A.) Uses of 98P4B6 Polynucleotides

II.A.1.) Monitoring of Genetic Abnormalities

The polynucleotides of the preceding paragraphs have a number of different specific uses. The human 98P4B6 gene maps to the chromosomal location set forth in the Example entitled “Chromosomal Mapping of 98P4B6.” For example, because the 98P4B6 gene maps to this chromosome, polynucleotides that encode different regions of the 98P4B6 proteins are used to characterize cytogenetic abnormalities of this chromosomal locale, such as abnormalities that are identified as being associated with various cancers. In certain genes, a variety of chromosomal abnormalities including rearrangements have been identified as frequent cytogenetic abnormalities in a number of different cancers (see e.g. Krajinovic et al., Mutat. Res. 382(3-4): 81-83 (1998); Johansson et al., Blood 86(10): 3905-3914 (1995) and Finger et al., P.N.A.S. 85(23): 9158-9162 (1988)). Thus, polynucleotides encoding specific regions of the 98P4B6 proteins provide new tools that can be used to delineate, with greater precision than previously possible, cytogenetic abnormalities in the chromosomal region that encodes 98P4B6 that may contribute to the malignant phenotype. In this context, these polynucleotides satisfy a need in the art for expanding the sensitivity of chromosomal screening in order to identify more subtle and less common chromosomal abnormalities (see e.g. Evans et al., Am. J. Obstet. Gynecol 171(4): 1055-1057 (1994)).

Furthermore, as 98P4B6 was shown to be highly expressed in prostate and other cancers, 98P4B6 polynucleotides are used in methods assessing the status of 98P486 gene products in normal versus cancerous tissues. Typically, polynucleotides that encode specific regions of the 98P4B6 proteins are used to assess the presence of perturbations (such as deletions, insertions, point mutations, or alterations resulting in a loss of an antigen etc.) in specific regions of the 98P4B6 gene, such as regions containing one or more motifs. Exemplary assays include both RT-PCR assays as well as single-strand conformation polymorphism (SSCP) analysis (see, e.g., Marrogi et al., J. Cutan. Pathol. 26(8): 369-378 (1999), both of which utilize polynucleotides encoding specific regions of a protein to examine these regions within the protein.

II.A.2.) Antisense Embodiments

Other specifically contemplated nucleic acid related embodiments of the invention disclosed herein are genomic DNA, cDNAs, ribozymes, and antisense molecules, as well as nucleic acid molecules based on an alternative backbone, or including alternative bases, whether derived from natural sources or synthesized, and include molecules capable of inhibiting the RNA or protein expression of 98P4B6. For example, antisense molecules can be RNAs or other molecules, including peptide nucleic acids (PNAs) or non-nucleic acid molecules such as phosphorothioate derivatives that specifically bind. DNA or RNA in a base pair-dependent manner. A skilled artisan can readily obtain these classes of nucleic acid molecules using the 98P4B6 polynucleotides and polynucleotide sequences disclosed herein.

Antisense technology entails the administration of exogenous oligonucleotides that bind to a target polynucleotide located within the cells. The term “antisense” refers to the fact that such oligonucleotides are complementary to their intracellular targets, e.g., 98P4B6. See for example, Jack Cohen, Oligodeoxynucleotides, Antisense Inhibitors of Gene Expression, CRC Press, 1989; and Synthesis 1:1-5 (1988). The 98P4B6 antisense oligonucleotides of the present invention include derivatives such as S-oligonucleotides (phosphorothioate derivatives or S-oligos, see, Jack Cohen, supra), which exhibit enhanced cancer cell growth inhibitory action. S-oligos (nucleoside phosphorothioates) are isoelectronic analogs of an oligonucleotide (O-oligo) in which a nonbridging oxygen atom of the phosphate group is replaced by a sulfur atom. The S-oligos of the present invention can be prepared by treatment of the corresponding O-oligos with 3H-1,2-benzodithiol-3-one-1,1-dioxide, which is a sulfur transfer reagent. See, e.g., Iyer, R. P. et al., J. Org. Chem. 55:4693-4698 (1990); and Iyer, R. P. et al., J. Am. Chem. Soc. 112:1253-1254 (1990). Additional 98P4B6 antisense oligonucleotides of the present invention include morpholino antisense oligonucleotides known in the art (see, e.g., Partridge et al., 1996, Antisense & Nucleic Acid Drug Development 6: 169-175).

The 98P4B6 antisense oligonucleotides of the present invention typically can be RNA or DNA that is complementary to and stably hybridizes with the first 100 5′ codons or last 100 3′ codons of a 98P4B6 genomic sequence or the corresponding mRNA. Absolute complementarity is not required, although high degrees of complementarity are preferred. Use of an oligonucleotide complementary to this region allows for the selective hybridization to 98P4B6 mRNA and not to mRNA specifying other regulatory subunits of protein kinase. In one embodiment, 98P4B6 antisense oligonucleotides of the present invention are 15 to 30-mer fragments of the antisense DNA molecule that have a sequence that hybridizes to 98P4B6 mRNA. Optionally, 98P4B6 antisense oligonucleotide is a 30-mer oligonucleotide that is complementary to a region in the first 10 5′ codons or last 10 3′ codons of 98P4B6. Alternatively, the antisense molecules are modified to employ ribozymes in the inhibition of 98P4B6 expression, see, e.g., L. A. Couture & D. T. Stinchcomb; Trends Genet. 12: 510-515 (1996).

II.A.3.) Primers and Primer Pairs

Further specific embodiments of these nucleotides of the invention include primers and primer pairs, which allow the specific amplification of polynucleotides of the invention or of any specific parts thereof, and probes that selectively or specifically hybridize to nucleic acid molecules of the invention or to any part thereof. Probes can be labeled with a detectable marker, such as, for example, a radioisotope, fluorescent compound, bioluminescent compound, a chemiluminescent compound, metal chelator or enzyme. Such probes and primers are used to detect the presence of a 98P4B6 polynucleotide in a sample and as a means for detecting a cell expressing a 98P4B6 protein.

Examples of such probes include polypeptides comprising all or part of the human 98P4B6 cDNA sequence shown in FIG. 2. Examples of primer pairs capable of specifically amplifying 98P4B6 mRNAs are also described in the Examples. As will be understood by the skilled artisan, a great many different primers and probes can be prepared based on the sequences provided herein and used effectively to amplify and/or detect a 98P4B6 mRNA.

The 98P4B6 polynucleotides of the invention are useful for a variety of purposes, including but not limited to their use as probes and primers for the amplification and/or detection of the 98P4B6 gene(s), mRNA(s), or fragments thereof; as reagents for the diagnosis and/or prognosis of prostate cancer and other cancers; as coding sequences capable of directing the expression of 98P4B6 polypeptides; as tools for modulating or inhibiting the expression of the 98P4B6 gene(s) and/or translation of the 98P4B6 transcript(s); and as therapeutic agents.

The present invention includes the use of any probe as described herein to identify and isolate a 98P4B6 or 98P4B6 related nucleic acid sequence from a naturally occurring source, such as humans or other mammals, as well as the isolated nucleic acid sequence per se, which would comprise all or most of the sequences found in the probe used.

II.A.4.) Isolation of 98P4B6-Encoding Nucleic Acid Molecules

The 98P4B6 cDNA sequences described, herein enable the isolation of other polynucleotides encoding 98P4B6 gene product(s), as well as the isolation of polynucleotides encoding 98P4B6 gene product homologs, alternatively spliced isoforms, allelic variants, and mutant forms of a 98P4B6 gene product as well as polynucleotides that encode analogs of 98P4B6-related proteins. Various molecular cloning methods that can be employed to isolate full length cDNAs encoding a 98P4B6 gene are well known (see, for example, Sambrook, J. et al., Molecular Cloning: A Laboratory Manual, 2d edition, Cold Spring Harbor Press, New York, 1989; Current Protocols in Molecular Biology. Ausubel et al., Eds., Wiley and Sons, 1995). For example, lambda phage cloning methodologies can be conveniently employed, using commercially available cloning systems (e.g., Lambda ZAP Express, Stratagene). Phage clones containing 98P4B6 gene cDNAs can be identified by probing with a labeled 98P4B6 cDNA or a fragment thereof. For example, in one embodiment, a 98P4B6 cDNA (e.g., FIG. 2) or a portion thereof can be synthesized and used as a probe to retrieve overlapping and full-length cDNAs corresponding to a 98P4B6 gene. A 98P4B6 gene itself can be isolated by screening genomic DNA libraries, bacterial artificial chromosome libraries (BACs), yeast artificial chromosome libraries (YACs), and the like, with 98P4B6 DNA probes or primers.

II.A.5.) Recombinant Nucleic Acid Molecules and Host-Vector Systems

The invention also provides recombinant DNA or RNA molecules containing a 98P4B6 polynucleotide, a fragment, analog or homologue thereof, including but not limited to phages, plasmids, phagemids, cosmids, YACs, BACs, as well as various viral and non-viral vectors well known in the art, and cells transformed or transfected with such recombinant DNA or RNA molecules. Methods for generating such molecules are well known (see, for example, Sambrook et al., 1989, supra).

The invention further provides a host-vector system comprising a recombinant DNA molecule containing a 98P4B6 polynucleotide, fragment, analog or homologue thereof within a suitable prokaryotic or eukaryotic host cell. Examples of suitable eukaryotic host cells include a yeast cell, a plant cell, or an animal cell, such as a mammalian cell or an insect cell (e.g., a baculovirus-infectible cell such as an Sf9 or HighFive cell). Examples of suitable mammalian cells include various prostate cancer cell lines such as DU145 and TsuPr1, other transfectable or transducible prostate cancer cell lines, primary cells (PrEC), as well as a number of mammalian cells routinely used for the expression of recombinant proteins (e.g., COS, CHO, 293, 293T cells). More particularly, a polynucleotide comprising the coding sequence of 98P4B6 or a fragment, analog or homolog thereof can be used to generate 98P4B6 proteins or fragments thereof using any number of host-vector systems routinely used and widely known in the art

A wide range of host-vector systems suitable for the expression of 98P4B6 proteins or fragments thereof are available, see for example, Sambrook et al., 1989, supra; Current Protocols in Molecular Biology, 1995, supra). Preferred vectors for mammalian expression include but are not limited to pcDNA 3.1 myc-His-tag (Invitrogen) and the retroviral vector pSRαtkneo (Muller et al., 1991, MCB 11:1785). Using these expression vectors, 98P4B6 can be expressed in several prostate cancer and non-prostate cell lines, including for example 293, 293T, rat-1, NIH 3T3 and TsuPr1. The host-vector systems of the invention are useful for the production of a 98P4B6 protein or fragment thereof. Such host-vector systems can be employed to study the functional properties of 98P4B6 and 98P4B6 mutations or analogs.

Recombinant human 98P4B6 protein or an analog or homolog or fragment thereof can be produced by mammalian cells transfected with a construct encoding a 98P4B6-related nucleotide. For example, 293T cells can be transfected with an expression plasmid encoding 98P4B6 or fragment, analog or homolog thereof, a 98P4B6-related protein is expressed in the 293T cells, and the recombinant 98P4B6 protein is isolated using standard purification methods (e.g., affinity purification using anti-98P4B6 antibodies). In another embodiment, a 98P4B6 coding sequence is subcloned into the retroviral vector pSRαMSVtkneo and used to infect various mammalian cell lines, such as NIH 3T3, TsuPr1, 293 and rat-1 in order to establish 98P4B6 expressing cell lines. Various other expression systems well known in the art can also be employed. Expression constructs encoding a leader peptide joined in frame to a 98P4B6 coding sequence can be used for the generation of a secreted form of recombinant 98P4B6 protein.

As discussed herein, redundancy in the genetic code permits variation in 98P4B6 gene sequences. In particular, it is known in the art that specific host species often have specific codon preferences, and thus one can adapt the disclosed sequence as preferred for a desired host. For example, preferred analog codon sequences typically have rare codons (i.e., codons having a usage frequency of less than about 20% in known sequences of the desired host) replaced with higher frequency codons. Codon preferences for a specific species are calculated, for example, by utilizing codon usage tables available on the INTERNET.

Additional sequence modifications are known to enhance protein expression in a cellular host. These include elimination of sequences encoding spurious polyadenylation signals, exon/intron splice site signals, transposon-like repeats, and/or other such well-characterized sequences that are deleterious to gene expression. The GC content of the sequence is adjusted to levels average for a given cellular host, as calculated by reference to known genes expressed in the host cell. Where possible, the sequence is modified to avoid predicted hairpin secondary mRNA structures. Other useful modifications include the addition of a translational initiation consensus sequence at the start of the open reading frame, as described in Kozak, Mol. Cell. Biol., 9:5073-5080 (1989). Skilled artisans understand that the general rule that eukaryotic ribosomes initiate translation exclusively at the 5′ proximal AUG codon is abrogated only under rare conditions (see, e.g., Kozak PNAS 92(7): 2662-2666, (1995) and Kozak NAR 15(20): 8125-8148 (1987)).

III.) 98P4B6-RELATED PROTEINS

Another aspect of the present invention provides 98P4B6-related proteins. Specific embodiments of 98P4B6 proteins comprise a polypeptide having all or part of the amino acid sequence of human 98P4B6 as shown in FIG. 2 or FIG. 3. Alternatively, embodiments of 98P4B6 proteins comprise variant, homolog or analog polypeptides that have alterations in the amino acid sequence of 98P4B6 shown in FIG. 2 or FIG. 3.

Embodiments of a 98P4B6 polypeptide include: a 98P4B6 polypeptide having a sequence shown in FIG. 2, a peptide sequence of a 98P4B6 as shown in FIG. 2 wherein T is U; at least 10 contiguous nucleotides of a polypeptide having the sequence as shown in FIG. 2; or, at least 10 contiguous peptides of a polypeptide having the sequence as shown in FIG. 2 where T is U. For example, embodiments of 98P4B6 peptides comprise, without limitation:

-   -   (I) a protein comprising, consisting essentially of, or         consisting of an amino acid sequence as shown in FIG. 2A-AL or         FIG. 3A-J;     -   (II) a 98P4B6-related protein that is at least 90, 91, 92, 93,         94, 95, 96, 97, 98, 99 or 100% homologous to an entire amino         acid sequence shown in FIG. 2A-AL;     -   (III) a 98P4B6-related protein that is at least 90, 91, 92, 93,         94, 95, 96, 97, 98, 99 or 100% identical to an entire amino acid         sequence shown in FIG. 2A-AL or 3A-J;     -   (IV) a protein that comprises at least one peptide set forth in         Tables VIII to XLIX, optionally with a proviso that it is not an         entire protein of FIG. 2;     -   (V) a protein that comprises at least one peptide set forth in         Tables VIII-XXI, collectively, which peptide is also set forth         in Tables XXII to XLIX, collectively, optionally with a proviso         that it is not an entire protein of FIG. 2;     -   (VI) a protein that comprises at least two peptides selected         from the peptides set forth in Tables VIII-XLIX, optionally with         a proviso that it is not an entire protein of FIG. 2;     -   (VII) a protein that comprises at least two peptides selected         from the peptides set forth in Tables VIII to XLIX collectively,         with a proviso that the protein is not a contiguous sequence         from an amino acid sequence of FIG. 2;     -   (VIII) a protein that comprises at least one peptide selected         from the peptides set forth in Tables VIII-XXI; and at least one         peptide selected from the peptides set forth in Tables XXII to         XLIX, with a proviso that the protein is not a contiguous         sequence from an amino acid sequence of FIG. 2;     -   (IX) a polypeptide comprising at least 5, 6, 7, 8, 9, 10, 11,         12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,         28, 29, 30, 31, 32, 33, 34, 35 amino acids of a protein of FIG.         3A, 3B, 3C, 3D, 3E, 3F, 3G, 3H, 3I or 3J in any whole number         increment up to 454, 45, 419, 490, 576, 490, 454, 454, 576, or         490 respectively that includes at least 1, 2, 3, 4, 5, 6, 7, 8,         9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,         25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid         position(s) having a value greater than 0.5 in the         Hydrophilicity profile of FIG. 5;     -   (X) a polypeptide comprising at least 5, 6, 7, 8, 9, 10, 11, 12,         13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,         29, 30, 31, 32, 33, 34, 35 amino adds of a protein of FIG. 3A,         3B, 3C, 3D, 3E, 3F, 3G, 3H, 3I or 3J in any whole number         increment up to 454, 45, 419, 490, 576, 490, 454, 454, 576, or         490 respectively that includes at least at least 1, 2, 3, 4, 5,         6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,         23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid         position(s) having a value less than 0.5 in the Hydropathicity         profile of FIG. 6;     -   (XI) a polypeptide comprising at least 5, 6, 7, 8, 9, 10, 11,         12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,         28, 29, 30, 31, 32, 33, 34, 35 amino adds of a protein of FIG.         3A, 3B, 3C, 3D, 3E, 3F, 3G, 3H, 3I or 3J in any whole number         increment up to 454, 45, 419, 490, 576, 490, 454, 454, 576, or         490 respectively that includes at least at least 1, 2, 3, 4, 5,         6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,         23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid         position(s) having a value greater than 0.5 in the Percent         Accessible Residues, profile of FIG. 7;     -   (XII) a polypeptide comprising at least 5, 6, 7, 8, 9, 10, 11,         12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,         28, 29, 30, 31, 32, 33, 34, 35 amino acids of a protein of FIG.         3A, 3B, 3C, 3D, 3E, 3F, 3G, 3H, 3I or 3J in any whole number         increment up to 454, 45, 419, 490, 576, 490, 454, 454, 576, or         490 respectively that includes at least at least 1, 2, 3, 4, 5,         6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,         23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid         position(s) having a value greater than 0.5 in the Average         Flexibility profile of FIG. 8;     -   (XIII) a polypeptide comprising at least 5, 6, 7, 8, 9, 10, 11,         12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,         28, 29, 30, 31, 32, 33, 34, amino acids of a protein of FIG. 3A,         3B, 3C, 3D, 3E, 3F, 3G, 3H, 3I or 3J in any whole number         increment up to 454, 45, 419, 490, 576, 490, 454, 454, 576, or         490 respectively that includes at least at least 1, 2, 3, 4, 5,         6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,         23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid         position(s) having a value greater than 0.5 in the Beta-turn         profile of FIG. 9;     -   (XIV) a peptide that occurs at least twice in Tables VIII-XXI         and XXII to XLIX, collectively;     -   (XV) a peptide that occurs at least three times in Tables         VIII-XXI and XXII to XLIX, collectively;     -   (XVI) a peptide that occurs at least four times in Tables         VIII-XXI and XXII to XLIX, collectively;     -   (XVII) a peptide that occurs at least five times in Tables         VIII-XXI and XXII to XLIX, collectively;     -   (XVIII) a peptide that occurs at least once in Tables VIII-XXI,         and at least once in tables XXII to XLIX;     -   (XIX) a peptide that occurs at least once in Tables VIII-XXI,         and at least twice in tables XXII to XLIX;     -   (XX) a peptide that occurs at least twice in Tables VIII-XXI,         and at least once in tables XXII to XLIX;     -   (XXI) a peptide that occurs at least twice in Tables VII-XXI,         and at least twice in tables XXII to XLIX;     -   (XXII) a peptide which comprises one two, three, four, or five         of the following characteristics, or an oligonucleotide encoding         such peptide:         -   i) a region of at least 5 amino acids of a particular             peptide of FIG. 3, in any whole number increment up to the             full length of that protein in FIG. 3, that includes an             amino acid position having a value equal to or greater than             0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in             the Hydrophilicity profile of FIG. 5;         -   ii) a region of at least 5 amino adds of a particular             peptide of FIG. 3, in any whole number increment up to the             full length of that protein in FIG. 3, that includes an             amino acid position having a value equal to or less than             0.5, 0.4, 0.3, 0.2, 0.1, or having a value equal to 0.0, in             the Hydropathicity profile of FIG. 6;         -   iii) a region of at least 5 amino adds of a particular             peptide of FIG. 3, in any whole number increment up to the             full length of that protein in FIG. 3, that includes an             amino acid position having a value equal to or greater than             0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in             the Percent Accessible Residues profile of FIG. 7;         -   iv) a region of at least 5 amino adds of a particular             peptide of FIG. 3, in any whole number increment up to the             full length of that protein in FIG. 3, that includes an             amino acid position having a value equal to or greater than             0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in             the Average Flexibility profile of FIG. 8; or,     -   v) a region of at least 5 amino acids of a particular peptide of         FIG. 3, in any whole number increment up to the full length of         that protein in FIG. 3, that includes an amino acid position         having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9,         or having a value equal to 1.0, in the Beta-turn profile of FIG.         9;     -   (XXIII) a composition comprising a peptide of (I)-(XXII) or an         antibody or binding region thereof together with a         pharmaceutical excipient and/or in a human unit dose form.     -   (XXIV) a method of using a peptide of (I)-(XXII), or an antibody         or binding region thereof or a composition of (XXIII) in a         method to modulate a cell expressing 98P4B6,     -   (XXV) a method of using a peptide of (I)-(XXII) or an antibody         or binding region thereof or a composition of (XXIII) in a         method to diagnose, prophylax, prognose, or treat an individual         who bears a cell expressing 98P4B6     -   (XXVI) a method of using a peptide of (I)-(XXII) or an antibody         or binding region thereof or a composition (XXIII) in a method         to diagnose, prophylax, prbgnose, or treat an individual who         bears a cell expressing 98P4B6, said cell from a cancer of a         tissue listed in Table I;     -   (XXVII) a method of using a peptide of (I)-(XXII) or an antibody         or binding region thereof or a composition of (XXIII) in a         method to diagnose, prophylax, prognose, or treat a cancer;     -   (XXVIII) a method of using a peptide of (I)-(XXII) or an         antibody or binding region thereof or a composition of (XXIII)         in a method to diagnose, prophylax, prognose, or treat a a         cancer of a tissue listed in Table I; and,     -   (XXIX) a method of using a peptide of (I)-(XXII) or an antibody         or binding region thereof or a composition     -   (XXIII) in a method to identify or characterize a modulator of a         cell expressing 98P4B6.

As used herein, a range is understood to specifically disclose all whole unit positions thereof.

Typical embodiments of the invention disclosed herein include 98P4B6 polynucleotides that encode specific portions of 98P4B6 mRNA sequences (and those which are complementary to such sequences) such as those that encode the proteins and/or fragments thereof, for example:

(a) 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 225, 250, 275, 300, 325, 350, 375, 400, 410, 420, 430, 440, 450, or 454 more contiguous amino adds of 98P4B6 variant 1; the maximal lengths relevant for other variants are: variant 52, 45 amino acids; variant 5, 419 amino adds, variant 6, 490, variant 7, 576 amino acids, variant 8, 490 amino adds, variant 13, 454, variant 14, 454 amino acids, variant 21, 576 amino adds, and variant 25, 490 amino adds.

In general, naturally occurring allelic variants of human 98P4B6 share a high degree of structural identity and homology (e.g., 90% or more homology). Typically, allelic variants of a 98P4B6 protein contain conservative amino acid substitutions within the 98P4B6 sequences described herein or contain a substitution of an amino acid from a corresponding position in a homologue of 98P4B6. One class of 98P4B6 allelic variants are proteins that share a high degree of homology with at least a small region of a particular 98P4B6 amino acid sequence, but further contain a radical departure from the sequence, such as a non-conservative substitution, truncation, insertion or frame shift. In comparisons of protein sequences, the terms, similarity, identity, and homology each have a distinct meaning as appreciated in the field of genetics. Moreover, orthology and paralogy can be important concepts describing the relationship of members of a given protein-family in one organism to the members of the same family in other organisms:

Amino acid abbreviations are provided in Table II. Conservative amino acid substitutions can frequently be made in a protein without altering either the conformation or the function of the protein. Proteins of the invention can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 conservative substitutions. Such changes include substituting any of isoleucine (I), valine (V), and leucine (L) for any other of these hydrophobic amino acids; aspartic acid (D) for glutamic acid (E) and vice versa; glutamine (Q) for asparagine (N) and vice versa; and serine (S) for threonine (T) and vice versa. Other substitutions can also be considered conservative, depending on the environment of the particular amino acid and its role in the three-dimensional structure of the protein. For example, glycine (G) and alanine (A) can frequently be interchangeable, as can alanine (A) and valine (V). Methionine (M), which is relatively hydrophobic, can frequently be interchanged with leucine and isoleucine, and sometimes with valine. Lysine (K) and arginine (R) are frequently interchangeable in locations in which the significant feature of the amino acid residue is its charge and the differing pK's of these two amino acid residues are not significant. Still other changes can be considered “conservative” in particular environments (see, e.g. Table III herein; pages 13-15 “Biochemistry” 2^(nd) ED. LubertStyered (Stanford University); Henikoff et al., PNAS 1992 Vol 89 10915-10919; Lei et al., J Biol Chem 1995 May 19; 270(20):11882-6).

Embodiments of the invention disclosed herein include a wide variety of art-accepted variants or analogs of 98P4B6 proteins such as polypeptides having amino acid insertions, deletions and substitutions. 98P4B6 variants can be made using methods known in the art such as site-directed mutagenesis, alanine scanning, and PCR mutagenesis. Site-directed mutagenesis (Carter et al., Nucl. Acids Res., 13:4331 (1986); Zoller et al., Nucl. Acids Res., 10:6487 (1987)), cassette mutagenesis (Wells et al., Gene, 34:315 (1985)), restriction selection mutagenesis (Wells et al., Philos. Trans. R. Soc. London SerA, 317:415 (1986)) or other known techniques can be performed on the cloned DNA to produce the 98P486 variant DNA.

Scanning amino acid analysis can also be employed to identify one or more amino adds along a contiguous sequence that is involved in a specific biological activity such as a protein-protein interaction. Among the preferred scanning amino adds are relatively small, neutral amino acids. Such amino acids include alanine, glycine, serine, and cysteine. Alanine is typically a preferred scanning amino acid among this group because it eliminates the side-chain beyond the beta-carbon and is less likely to alter the main-chain conformation of the variant. Alanine is also typically preferred because it is the most common amino acid. Further, it is frequently found in both buried and exposed positions (Creighton, The Proteins, (W.H. Freeman & Co., N.Y.); Chothia, J. Mol. Biol., 150:1 (1976)). If alanine substitution does not yield adequate amounts of variant, an isosteric amino acid can be used.

As defined herein, 98P4B6 variants, analogs or homologs, have the distinguishing attribute of having at least one epitope that is “cross reactive” with a 98P4B6 protein having an amino acid sequence of FIG. 3. As used in this sentence, “cross reactive” means that an antibody or T cell that specifically binds to a 98P4B6 variant also specifically binds to a 98P4B6 protein having an amino acid sequence set forth in FIG. 3. A polypeptide ceases to be a variant of a protein shown in FIG. 3, when it no longer contains any epitope capable of being recognized by an antibody or T cell that specifically binds to the starting 98P4B6 protein. Those skilled in the art understand that antibodies that recognize proteins bind to epitopes of varying size, and a grouping of the order of about four or five amino acids, contiguous or not, is regarded as a typical number of amino acids in a minimal epitope. See, e.g., Nair et al., J. Immunol. 2000 165(12): 6949-6955; Hebbes et al., Mol Immunol (1989) 26(9):865-73; Schwartz et al., J Immunol (1985) 135(4):2598-608.

Other classes of 98P4B6-related protein variants share 70%, 75%, 80%, 85% or 90% or more similarity with an amino acid sequence of FIG. 3, or a fragment thereof. Another specific class of 98P4B6 protein variants or analogs comprises one or more of the 98P4B6 biological motifs described herein or presently known in the art. Thus, encompassed by the present invention are analogs of 98P4B6 fragments (nucleic or amino acid) that have altered functional (e.g. immunogenic) properties relative to the starting fragment. It is to be appreciated that motifs now or which become part of the art are to be applied to the nucleic or amino acid sequences of FIG. 2 or FIG. 3.

As discussed herein, embodiments of the claimed invention include polypeptides containing less than the full amino acid sequence of a 98P4B6 protein shown in FIG. 2 or FIG. 3. For example, representative embodiments of the invention comprise peptides/proteins having any 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more contiguous amino acids of a 98P4B6 protein shown in FIG. 2 or FIG. 3.

Moreover, representative embodiments of the invention disclosed herein include polypeptides consisting of about amino acid 1 to about amino acid 10 of a 98P4B6 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 10 to about amino acid 20 of a 98P4B6 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 20 to about amino acid 30 of a 98P4B6 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 30 to about amino acid 40 of a 98P4B6 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 40 to about amino acid 50 of a 98P4B6 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 50 to about amino acid 60 of a 98P4B6 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 60 to about amino acid 70 of a 98P4B6 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 70 to about amino acid 80 of a 98P4B6 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 80 to about amino acid 90 of a 98P4B6 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 90 to about amino acid 100 of a 98P4B6 protein shown in FIG. 2 or FIG. 3, etc. throughout the entirety of a 98P4B6 amino acid sequence. Moreover, polypeptides consisting of about amino acid 1 (or 20 or 30 or 40 etc.) to about amino acid 20, (or 130, or 140 or 150 etc.) of a 98P4B6 protein shown in FIG. 2 or FIG. 3 are embodiments of the invention. It is to be appreciated that the starting and stopping positions in this paragraph refer to the specified position as well as that position plus or minus 5 residues. 98P4B6-related proteins are generated using standard peptide synthesis technology or using chemical cleavage methods well known in the art. Alternatively, recombinant methods can be used to generate nucleic acid molecules that encode a 98P4B6-related protein. In one embodiment, nucleic acid molecules provide a means to generate defined fragments of a 98P4B6 protein (or variants, homologs or analogs thereof).

III.A.) Motif-Bearing Protein Embodiments

Additional illustrative embodiments of the invention disclosed herein include 98P4B6 polypeptides comprising the amino acid residues of one or more of the biological motifs contained within a 98P4B6 polypeptide sequence set forth in FIG. 2 or FIG. 3. Various motifs are known in the art, and a protein can be evaluated for the presence of such motifs by a number of publidy available sequence analysis tools (see, e.g., PFAM; BCM Search Launcher; PSORT; CBS; InterProScan; ScanProsite; Epimatrix™ and Epimer™, Brown University; and BIMAS).

Motif bearing subsequences of all 98P486 variant proteins are set forth and identified in Tables VIII-XXI and XXII-XLIX.

Table V sets forth several frequently occurring motifs based on pfam searches. The columns of Table V list (1) motif name abbreviation, (2) percent identity found amongst the different member of the motif family, (3) motif name or description and (4) most common function; location information is included if the motif is relevant for location.

Polypeptides comprising one or more of the 98P4B6 motifs discussed above are useful in elucidating the specific characteristics of a malignant phenotype in view of the observation that the 98P4B6 motifs discussed above are associated with growth dysregulation and because 98P4B6 is overexpressed in certain cancers (See, e.g., Table I). Casein kinase II, cAMP and camp-dependent protein kinase, and Protein Kinase C, for example, are enzymes known to be associated with the development of the malignant phenotype (see e.g. Chen et al., Lab Invest., 78(2): 165-174 (1998); Gaiddon et al., Endocrinology 136(10): 4331-4338 (1995); Hall et al., Nucleic Adds Research 24(6): 1119-1126 (1996); Peterziel et al., Oncogene 18(46): 6322-6329 (1999) and O'Brian, Oncol. Rep. 5(2): 305-309 (1998)). Moreover, both glycosylation and myristoylaion are protein modifications also associated with cancer and cancer progression (see e.g. Dennis et al., Biochem. Biophys. Acta 1473(1):21-34 (1999); Raju et al., Exp. Cell Res. 235(1): 145-154 (1997)). Amidation is another protein modification also associated with cancer and cancer progression (see e.g. Treston et al., J. Natl. Cancer Inst. Monogr. (13): 169-175 (1992)).

In another embodiment, proteins of the invention comprise one or more of the immunoreactive epitopes identified in accordance with art-accepted methods, such as the peptides set forth in Tables VIII-XXI and XXII-XLIX. CTL epitopes can be determined using specific algorithms to identify peptides within a 98P4B6 protein that are capable of optimally binding to specified HLA alleles (e.g., Table IV; Epimatrix™ and Epimer™, Brown University; and BIMAS). Moreover, processes for identifying peptides that have sufficient binding affinity for HLA molecules and which are correlated with being immunogenic epitopes, are well known in the art, and are carried out without undue experimentation. In addition, processes for identifying peptides that are immunogenic epitopes, are well known in the art, and are carried out without undue experimentation either in vitro or in vivo.

Also known in the art are principles for creating analogs of such epitopes in order to modulate immunogenicity. For example, one begins with an epitope that bears a CTL or HTL motif (see, e.g., the HLA Class I and HLA Class II motifs/supermotifs of Table IV). The epitope is analoged by substituting out an amino acid at one of the specified positions, and replacing it with another amino acid specified for that position. For example, on the basis of residues defined in Table IV, one can substitute out a deleterious residue in favor of any other residue, such as a preferred residue; substitute a less-preferred residue with a preferred residue; or substitute an originally-occurring preferred residue with another preferred residue. Substitutions can occur at primary anchor positions or at other positions in a peptide; see, e.g., Table IV.

A variety of references reflect the art regarding the identification and generation of epitopes in a protein of interest as well as analogs thereof. See, for example, WO 97/33602 to Chesnut et al.; Sette, Immunogenetics 1999 50(3-4): 201-212; Selte et al., J. Immunol. 2001 166(2): 1389-1397; Sidney et al., Hum. Immunol. 199758(1): 12-20; Kondo et al., Immunogenetics 1997 45(4): 249-258; Sidney et al., J. Immunol. 1996 157(8): 3480-90; and Falk et al., Nature 351: 290-6 (1991); Hunt et al., Science 255:1261-3 (1992); Parker et al., J. Immunol. 149:3580-7 (1992); Parker et al., J. Immunol. 152:163-75 (1994)); Kast et al., 1994 152(8): 3904-12; Borras-Cuesta et al., Hum. Immunol. 2000 61(3): 266-278; Alexander et al., J. Immunol. 2000 164(3); 164(3): 1625-1633; Alexander et al., PMID: 7895164, UI: 95202582; O'Sullivan et al., J. Immunol. 1991 147(8): 2663-2669; Alexander et al., Immunity 1994 1(9): 751-761 and Alexander et al., Immunol. Res. 1998 18(2): 79-92.

Related embodiments of the invention include polypeptides comprising combinations of the different motifs set forth in Table VI, and/or, one or more of the predicted CTL epitopes of Tables VIII-XXI and XXII-XLIX, and/or, one or more of the predicted HTL epitopes of Tables XLVI-XLIX, and/or, one or more of the T cell binding motifs known in the art. Preferred embodiments contain no insertions, deletions or substitutions either within the motifs or within the intervening sequences of the polypeptides. In addition, embodiments which include a number of either N-terminal and/or C-terminal amino acid residues on either side of these motifs may be desirable (to, for example, include a greater portion of the polypeptide architecture in which the motif is located). Typically, the number of N-terminal and/or C-terminal amino acid residues on either side of a motif is between about 1 to about 100 amino acid residues, preferably 5 to about 50 amino acid residues. 98P4B6-related proteins are embodied in many forms, preferably in isolated form. A purified 98P4B6 protein molecule will be substantially free of other proteins or molecules that impair the binding of 98P4B6 to antibody, T cell or other ligand. The nature and degree of isolation and purification will depend on the intended use. Embodiments of a 98P4B6-related proteins include purified 98P4B6-related proteins and functional, soluble 98P4B6-related proteins. In one embodiment, a functional, soluble 98P4B6 protein or fragment thereof retains the ability to be bound by antibody, T cell or other ligand.

The invention also provides 98P4B6 proteins comprising biologically active fragments of a 98P4B6 amino acid sequence shown in FIG. 2 or FIG. 3. Such proteins exhibit properties of the starting 98P4B6 protein, such as the ability to elicit the generation of antibodies that specifically bind an epitope associated with the starting 98P4B6 protein; to be bound by such antibodies; to elicit the activation of HTL or CTL; and/or, to be recognized by HTL or CTL that also specifically bind to the starting protein.

98P4B6-related polypeptides that contain particularly interesting structures can be predicted and/or identified using various analytical techniques well known in the art including, for example, the methods of Chou-Fasman, Garnier-Robson, Kyle-Doolittle, Eisenberg, Karplus-Schultz or Jameson-Wolf analysis, or based on immunogenidty. Fragments that contain such structures are particularly useful in generating subunit-specific anti-98P4B6 antibodies or T cells or in identifying cellular factors that bind to 98P4B6. For example, hydrophilicity profiles can be generated, and immunogenic peptide fragments identified, using the method of Hopp, T. P. and Woods, K. R., 1981, Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828. Hydropathidty profiles can be generated, and immunogenic peptide fragments identified, using the method of Kyte, J. and Doolittle, R. F., 1982, J. Mol. Biol. 157:105-132. Percent (%) Accessible Residues profiles can be generated, and immunogenic peptide fragments identified, using the method of Janin J., 1979, Nature 277:491-492. Average Flexibility profiles can be generated, and immunogenic peptide fragments identified, using the method of Bhaskaran R., Ponnuswamy P. K., 1988, Int. J. Pept. Protein Res. 32:242-255. Beta-turn profiles can be generated, and immunogenic peptide fragments identified, using the method of Deleage, G., Roux B., 1987, Protein Engineering 1:289-294.

CTL epitopes can be determined using specific algorithms to identify peptides within a 98P4B6 protein that are capable of optimally binding to specified HLA alleles (e.g., by using the SYFPEITHI site; the listings in Table IV(A)E); Epimatrix™ and Epimer™, Brown University; and BIMAS. Illustrating this, peptide epitopes from 98P4B6 that are presented in the context of human MHC Class I molecules, e.g., HLA-A1, A2, A3, A11, A24, B7 and B35 were predicted (see, e.g., Tables VIII-XXI, XXII-XLIX). Specifically, the complete amino acid sequence of the 98P4B6 protein and relevant portions of other variants, i.e., for HLA Class I predictions 9 flanking residues on either side of a point mutation or exon junction, and for HLA Class II predictions 14 flanking residues on either side of a point mutation or exon junction corresponding to that variant, were entered into the HLA Peptide Motif Search algorithm found in the Bioinformatics and Molecular Analysis Section (BIMAS) web site listed above; in addition to the site SYFPEITHI.

The HLA peptide motif search algorithm was developed by Dr. Ken Parker based on binding of specific peptide sequences in the groove of HLA Class I molecules, in particular HLA-A2 (see, e.g., Falk et al, Nature 351: 2904 (1991); Hunt et at, Science 255:1261-3 (1992); Parker et at, J. Immunol. 149:3580-7 (1992); Parker et at, J. Immunol. 152:163-75 (1994)). This algorithm allows location and ranking of 8-mer, 9-mer, and 10mer peptides from a complete protein sequence for predicted binding to HLA-A2 as well as numerous other HLA Class I molecules. Many HLA class I binding peptides are 8-, 9-, 10 or 11-mers. For example, for Class I HLA-A2, the epitopes preferably contain a leucine (L) or methionine (M) at position 2 and a valine (V) or leucine (L) at the C-terminus (see, e.g., Parker et al., J. Immunol. 149:3580-7 (1992)). Selected results of 98P4B6 predicted binding peptides are shown in Tables VIII-XXI and XXII-XLIX herein. In Tables VIII-XXI and XXII-XLVII, selected candidates, 9-mers and 10-mers, for each family member are shown along with their location, the amino acid sequence of each specific peptide, and an estimated binding score. In Tables XLVI-XLIX, selected candidates, 15-mers, for each family member are shown along with their location, the amino acid sequence of each specific peptide, and an estimated binding score. The binding score corresponds to the estimated half time of dissociation of complexes containing the peptide at 37° C. at pH 6.5. Peptides with the highest binding score are predicted to be the most tightly bound to HLA Class I on the cell surface for the greatest period of time and thus represent the best immunogenic targets for T-cell recognition.

Actual binding of peptides to an HLA allele can be evaluated by stabilization of HLA expression on the antigen-processing defective cell line T2 (see, e.g., Xue et al., Prostate 30:73-8 (1997) and Peshwa et al., Prostate 36:129-38 (1998)). Immunogenicity of specific peptides can be evaluated in vmro by sbtmulaion of CD8+ cytotoxic T lymphocytes (CTL) in the presence of antigen presenting cells such as dendritic cells.

It is to be appreciated that every epitope predicted by the BIMAS site, Epimer™ and Epimatrix™ sites, or specified by the HLA class I or class II motifs available in the art or which become part of the art such as set forth in Table IV (or determined using SYFPEITHI or BIMAS) are to be “applied” to a 98P4B6 protein in accordance with the invention. As used in this context “applied” means that a 98P4B6 protein is evaluated, e.g., visually or by computer-based patterns finding methods, as appreciated by those of skill in the relevant art. Every subsequence of a 98P4B6 protein of 8, 9, 10, or 11 amino acid residues that bears an HLA Class I motif, or a subsequence of 9 or more amino acid residues that bear an HLA Class II motif are within the scope of the invention.

III.B.) Expression of 98P4B6-Related Proteins

In an embodiment described in the examples that follow, 98P4B6 can be conveniently expressed in cells (such as 293T cells) transfected with a commercially available expression vector such as a CMV-driven expression vector encoding 98P4B6 with a C-terminal 6× His and MYC tag (pcDNA3.1/mycHIS, Invitrogen or Tag5, GenHunter Corporation, Nashville Tenn.). The Tag5 vector provides an IgGK secretion signal that can be used to facilitate the production of a secreted 98P4B6 protein in transfected cells. The secreted HIS-tagged 98P4B6 in the culture media can be purified, e.g., using a nickel column using standard techniques.

III.C.) Modifications of 98P4B6-Related Proteins

Modifications of 98P4B6-related proteins such as covalent modifications are included within the scope of this invention. One type of covalent modification includes reacting targeted amino acid residues of a 98P4B6 polypeptide with an organic derivatizing agent that is capable of reacting with selected side chains or the N- or C-terminal residues of a 98P4B6 protein. Another type of covalent modification of a 98P4B6 polypeptide included within the scope of this invention comprises altering the native glycosylation pattern of a protein of the invention. Another type of covalent modification of 98P4B6 comprises linking a 98P4B6 polypeptide to one of a variety of nonproteinaceous polymers, e.g., polyethylene glycol (PEG), polypropylene glycol, or polyoxyalkylenes, in the manner set forth in U.S. Pat. No. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192 or 4,179,337.

The 98P4B6-related proteins of the present invention can also be modified to form a chimeric molecule comprising 98P4B6 fused to another, heterologous polypeptide or amino acid sequence. Such a chimeric molecule can be synthesized chemically or recombinantly. A chimeric molecule can have a protein of the invention fused to another tumor-associated antigen or fragment thereof. Alternatively, a protein in accordance with the invention can comprise a fusion of fragments of a 98P4B6 sequence (amino or nucleic acid) such that a molecule is created that is not, through its length, directly homologous to the amino or nucleic acid sequences shown in FIG. 2 or FIG. 3. Such a chimeric molecule can comprise multiples of the same subsequence of 98P4B6. A chimeric molecule can comprise a fusion of a 98P4B6-related protein with a polyhistidine epitope tag, which provides an epitope to which immobilized nickel can selectively bind, with cytokines or with growth factors. The epitope tag is generally placed at the amino- or carboxyl-terminus of a 98P4B6 protein. In an alternative embodiment, the chimeric molecule can comprise a fusion of a 98P4B6-related protein with an immunoglobulin or a particular region of an immunoglobulin. For a bivalent form of the chimeric molecule (also referred to as an “immunoadhesin”), such a fusion could be to the Fc region of an IgG molecule. The Ig fusions preferably include the substitution of a soluble (transmembrane domain deleted or inactivated) form of a 98P4B6 polypeptide in place of at least one variable region within an Ig molecule. In a preferred embodiment, the immunoglobulin fusion includes the hinge, CH2 and CH3, or the hinge, CHI, CH2 and CH3 regions of an IgGI molecule. For the production of immunoglobulin fusions see, e.g., U.S. Pat. No. 5,428,130 issued Jun. 27, 1995.

III.D.) Uses of 98P4B6-Related Proteins

The proteins of the invention have a number of different specific uses. As 98P4B6 is highly expressed in prostate and other cancers, 98P4B6-related proteins are used in methods that assess the status of 98P4B6 gene products in normal versus cancerous tissues, thereby elucidating the malignant phenotype. Typically, polypeptides from specific regions of a 98P4B6 protein are used to assess the presence of perturbations (such as deletions, insertions, point mutations etc.) in those regions (such as regions containing one or more motifs). Exemplary assays utilize antibodies or T cells targeting 98P4B6-related proteins comprising the amino acid residues of one or more of the biological motifs contained within a 98P4B6 polypeptide sequence in order to evaluate the characteristics of this region in normal versus cancerous tissues or to elicit an immune response to the epitope. Alternatively, 98P4B6-related proteins that contain the amino acid residues of one or more of the biological motifs in a 98P4B6 protein are used to screen for factors that interact with that region of 98P4B6.

98P4B6 protein fragments/subsequences are particularly useful in generating and characterizing domain-specific antibodies (e.g., antibodies recognizing an extracellular or intracellular epitope of a 98P4B6 protein), for identifying agents or cellular factors that bind to 98P4B6 or a particular structural domain thereof, and in various therapeutic and diagnostic contexts, including but not limited to diagnostic assays, cancer vaccines and methods of preparing such vaccines.

Proteins encoded by the 98P4B6 genes, or by analogs, homologs or fragments thereof, have a variety of uses, including but not limited to generating antibodies and in methods for identifying ligands and other agents and cellular constituents that bind to a 98P4B6 gene product. Antibodies raised against a 98P4B6 protein or fragment thereof are useful in diagnostic and prognostic assays, and imaging methodologies in the management of human cancers characterized by expression of 98P4B6 protein, such as those listed in Table I. Such antibodies can be expressed intracellularly and used in methods of treating patients with such cancers. 98P4B6-related nucleic adds or proteins are also used in generating HTL or CTL responses.

Various immunological assays useful for the detection of 98P4B6 proteins are used, including but not limited to various types of radioimmunoassays, enzyme inked immunosorbent assays (ELISA), enzyme-linked immunofluorescent assays (ELIFA), immunocytochemical methods, and the like. Antibodies can be labeled and used as immunological imaging reagents capable of detecting 98P4B6-expressing cells (e.g., in radioscintigraphic imaging methods). 98P486 proteins are also particularly useful in generating cancer vaccines, as further described herein.

IV.) 98P4B6 Antibodies Another aspect of the invention provides antibodies that bind to 98P4B6-related proteins. Preferred antibodies specifically bind to a 98P4B6-related protein and do not bind (or bind weakly) to peptides or proteins that are not 98P4B6-related proteins under physiological conditions. In this context, examples of physiological conditions include: 1) phosphate buffered saline; 2) Tris-buffered saline containing 25 mM Tris and 150 mM NaCl; or normal saline (0.9% NaCl); 4) animal serum such as human serum; or, 5) a combination of any of 1) through 4); these reactions preferably taking place at pH 7.5, alternatively in a range of pH 7.0 to 8.0, or alternatively in a range of pH 6.5 to 8.5; also, these reactions taking place at a temperature between 4° C. to 37° C. For example, antibodies that bind 98P4B6 can bind 98P4B6-related proteins such as the homologs or analogs thereof.

98P4B6 antibodies of the invention are particularly useful in cancer (see, e.g., Table I) diagnostic and prognostic assays, and imaging methodologies. Similarly, such antibodies are useful in the treatment, diagnosis, and/or prognosis of other cancers, to the extent 98P4B6 is also expressed or overexpressed in these other cancers. Moreover, intracellularly expressed antibodies (e.g., single chain antibodies) are therapeutically useful in treating cancers in which the expression of 98P4B6 is involved, such as advanced or metastatic prostate cancers.

The invention also provides various immunological assays useful for the detection and quantification of 98P4B6 and mutant 98P4B6-related proteins. Such assays can comprise one or more 98P4B6 antibodies capable of recognizing and binding a 98P4B6-related protein, as appropriate. These assays are performed within various immunological assay formats well known in the art, including but not limited to various types of radioimmunoassays, enzyme-linked immunosorbent assays (ELISA), enzyme-linked immunofluorescent assays (ELIFA), and the like.

Immunological non-antibody assays of the invention also comprise T cell immunogenicity assays (inhibitory or stimulatory) as well as major histocompatibility complex (MHC) binding assays.

In addition, immunological imaging methods capable of detecting prostate cancer and other cancers expressing 98P4B6 are also provided by the invention, including but not limited to radioscintigraphic imaging methods using labeled 98P4B6 antibodies. Such assays are clinically useful in the detection, monitoring, and prognosis of 98P4B6 expressing cancers such as prostate cancer.

98P4B6 antibodies are also used in methods for purifying a 98P4B6-related protein and for isolating 98P4B6 homologues and related molecules. For example, a method of purifying a 98P4B6-related protein comprises incubating a 98P4B6 antibody, which has been coupled to a solid matrix, with a lysate or other solution containing a 98P4B6-related protein under conditions that permit the 98P4B6 antibody to bind to the 98P4B6-related protein; washing the solid matrix to eliminate impurities; and eluting the 98P4B6-related protein from the coupled antibody. Other uses of 98P4B6 antibodies in accordance with the invention include generating anti-idiotypic antibodies that mimic a 98P4B6 protein.

Various methods for the preparation of antibodies are well known in the arL For example, antibodies can be prepared by immunizing a suitable mammalian host using a 98P4B6-related protein, peptide, or fragment, in isolated or immunoconjugated form (Antibodies: A Laboratory Manual, CSH Press, Eds., Harlow, and Lane (1988); Harlow, Antibodies, Cold Spring Harbor Press, NY (1989)). In addition, fusion proteins of 98P4B6 can also be used, such as a 98P4B6 GST-fusion protein. In a particular embodiment, a GST fusion protein comprising all or most of the amino acid sequence of FIG. 2 or FIG. 3 is produced, then used as an immunogen to generate appropriate antibodies. In another embodiment, a 98P4B6-related protein is synthesized and used as an immunogen.

In addition, naked DNA immunization techniques known in the art are used (with or without purified 98P4B6-related protein or 98P486 expressing cells) to generate an immune response to the encoded immunogen (for review, see Donnelly et al, 1997, Ann. Rev. Immunol. 15:617-648).

The amino acid sequence of a 98P4B6 protein as shown in FIG. 2 or FIG. 3 can be analyzed to select specific regions of the 98P4B6 protein for generating antibodies. For example, hydrophobicity and hydrophilicity analyses of a 98P4B6 amino acid sequence are used to identify hydrophilic regions in the 98P4B6 structure. Regions of a 98P4B6 protein that show immunogenic structure, as well as other regions and domains, can readily be identified using various other methods known in the ark such as Chou-Fasman, Garnier-Robson, Kyte-Doolittle, Eisenberg, Karplus-Schultz or Jameson-Woff analysis. Hydrophilicity profiles can be generated using the method of Hopp, T. P. and Woods, K. R., 1981, Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828. Hydropathicity profiles can be generated using the method of Kyte, J. and Doolittle, R. F., 1982, J. Mol. Biol. 157:105-132. Percent (%) Accessible Residues profiles can be generated using the method of Janin J., 1979, Nature 277:491-492. Average Flexibility profiles can be generated using the method of Bhaskaran R., Ponnuswamy P. K., 1988, Int. J. Pept. Protein Res. 32:242-255. Beta-turn profiles can be generated using the method of Deleage, G., Roux B., 1987, Protein Engineering 1:289-294. Thus, each region identified by any of these programs or methods is within the scope of the present invention. Methods for the generation of 98P4B6 antibodies are further illustrated by way of the examples provided herein. Methods for preparing a protein or polypeptide for use as an immunogen are well known in the art. Also well known in the art are methods for preparing immunogenic conjugates of a protein with a carrier, such as BSA, KLH or other carrier protein. In some circumstances, direct conjugation using, for example, carbodiimide reagents are used; in other instances linking reagents such as those supplied by Pierce Chemical Co., Rockford, Ill., are effective. Administration of a 98P4B6 immunogen is often conducted by injection over a suitable time period and with use of a suitable adjuvant, as is understood in the art. During the immunization schedule, titers of antibodies can be taken to determine adequacy of antibody formation. 98P4B6 monoclonal antibodies can be produced by various means well known in the art. For example, immortalized cell lines that secrete a desired monoclonal antibody are prepared using the standard hybridoma technology of Kohler and Milstein or modifications that immortalize antibody-producing B cells, as is generally known. Immortalized cell lines that secrete the desired antibodies are screened by immunoassay in which the antigen is a 98P4B6-related protein. When the appropriate immortalized cell culture is identified, the cells can be expanded and antibodies produced either from in vitro cultures or from ascites fluid.

The antibodies or fragments of the invention can also be produced, by recombinant means. Regions that bind specifically to the desired regions of a 98P4B6 protein can also be produced in the context of chimeric or complementarity-determining region (CDR) grafted antibodies of multiple species origin. Humanized or human 98P4B6 antibodies can also be produced, and are preferred for use in therapeutic contexts. Methods for humanizing murine and other non-human antibodies, by substituting one or more of the non-human antibody CDRs for corresponding human antibody sequences, are well known (see for example, Jones et al., 1986, Nature 321: 522-525; Riechmann et al., 1988, Nature 332: 323-327; Verhoeyen et al., 1988, Science 239: 1534-1536). See also, Carter et al., 1993, Proc. Nat. Acad. Sci. USA 89: 4285 and Sims et al., 1993, J. Immunol. 151: 2296.

Methods for producing fully human monoclonal antibodies include phage display and transgenic methods (for review, see Vaughan et al., 1998, Nature Biotechnology 16: 535-539). Fully human 98P4B6 monoclonal antibodies can be generated using cloning technologies employing large human Ig gene combinatorial libraries (i.e., phage display) (Griffiths and Hoogenboom, Building an in vitro immune system: human antibodies from phage display libraries. In: Protein Engineering of Antibody Molecules for Prophylactic and Therapeutic Applications in Man, Clark, M. (Ed.), Nottingham Academic, pp 4564 (1993); Burton and Barbas, Human Antibodies from combinatorial libraries. Id., pp 65-82). Fully human 98P4B6 monoclonal antibodies can also be produced using transgenic mice engineered to contain human immunoglobulin gene loci as described in PCT Patent Application WO98/24893, Kucherlapati and Jakobovits et al., published Dec. 3, 1997 (see also, Jakobovits, 1998, Exp. Opin. Invest Drugs 7(4): 607-614; U.S. Pat. Nos. 6,162,963 issued 19 Dec. 2000; 6,150,584 issued 12 Nov. 2000; and, 6,114,598 issued 5 Sep. 2000). This method avoids the in vitro manipulation required with phage display technology and efficiently produces high affinity authentic human antibodies.

Reactivity of 98P4B6 antibodies with a 98P4B6-related protein can be established by a number of well known means, including Western blot, immunoprecipitation, ELISA, and FACS analyses using, as appropriate, 98P4B6-related proteins, 98P4B6-expressing cells or extracts thereof. A 98P4B6 antibody or fragment thereof can be labeled with a detectable marker or conjugated to a second molecule. Suitable detectable markers include, but are not limited to, a radioisotope, a fluorescent compound, a bioluminescent compound, chemiluminescent compound, a metal chelator or an enzyme. Further, bi-specific antibodies specific for two or more 98P4B6 epitopes are generated using methods generally known in the art. Homodimeric antibodies can also be generated by cross-linking techniques known in the art (e.g., Wolff et al., Cancer Res. 53: 2560-2565).

V.) 98P4B6 CELLULAR IMMUNE RESPONSES

The mechanism by which T cells recognize antigens has been delineated. Efficacious peptide epitope vaccine compositions of the invention induce a therapeutic or prophylactic immune responses in very broad segments of the world-wide population. For an understanding of the value and efficacy of compositions of the invention that induce cellular immune responses, a brief review of immunology-related technology is provided.

A complex of an HLA molecule and a peptidic antigen acts as the ligand recognized by HLA-restricted T cells (Buus, S. et al., Cell 47:1071, 1986; Babbitt, B. P. et al., Nature 317:359, 1985; Townsend, A. and Bodmer, H., Annu. Rev. Immunol. 7:601, 1989; Germain, R. N., Annu. Rev. Immunol. 11:403, 1993). Through the study of single amino acid substituted antigen analogs and the sequencing of endogenously bound, naturally processed peptides, critical residues that correspond to motifs required for specific binding to HLA antigen molecules have been identified and are set forth in Table IV (see also, e.g., Southwood, et al., J. Immunol. 160:3363, 1998; Rammensee, et al., Immunogenetics 41:178, 1995; Rammensee et al., SYFPEITHI; Sette, A. and Sidney, J. Curr. Opin. Immunol 10:478, 1998; Engelhard, V. H., Curr. Opin. Immunol. 6:13, 1994; Selle, A. and Grey, H. M. Curr. Opin. Immunol. 4:79, 1992; Sinigaglia, F. and Hammer, J. Curr. Biol. 6:52, 1994; Ruppert et al., Cell 74:929-937, 1993; Kondo et al., J. Immunol. 155:4307-4312, 1995; Sidney et al., J. Immunol. 157:3480-3490, 1996; Sidney et al., Human Immunol. 45:79-93, 1996; Sette, A. and Sidney, J. Immunogenetics 1999 November; 50(3-4):201-12, Review).

Furthermore, x-ray crystallographic analyses of HLA-peptide complexes have revealed pockets within the peptide binding cleft/groove of HLA molecules which accommodate, in an allele-specific mode, residues borne by peptide ligands; these residues in turn determine the HLA binding capacity of the peptides in which they are present. (See, e.g., Madden, D. R. Annu. Rev. Immunol. 13:587, 1995; Smith, et al., Immunity 4:203, 1996; Fremont et al., Immunity 8:305, 1998; Stem et al., Structure 2:245, 1994; Jones, E. Y. Curr. Opin. Immunol 9:75, 1997; Brown, J. H. et al., Nature 364:33, 1993; Guo, H. C. et al., Proc. Natl. Acad. Sci. USA 90:8053, 1993; Guo, H. C. et al., Nature 360:364, 1992; Silver, M. L. et al., Nature 360:367, 1992; Matsumura, M. et al., Science 257:927, 1992; Madden et al., Cell 70:1035, 1992; Fremont, D. H. et al., Science 257:919, 1992; Saper, M. A., Bjorkman, P. J. and Wiley, D.C., J. Mol. Biol. 219:277, 1991.)

Accordingly, the definition of class I and class II allele-specific HLA binding motifs, or class I or class II supermotifs allows identification of regions within a protein that are correlated with binding to particular HLA antigen(s).

Thus, by a process of HLA motif identification, candidates for epitope-based vaccines have been identified; such candidates can be further evaluated by HLA-peptide binding assays to determine binding affinity and/or the time period of association of the epitope and its corresponding HLA molecule. Additional confirmatory work can be performed to select, amongst these vaccine candidates, epitopes with preferred characteristics in terms of population coverage, and/or immunogenicity.

Various strategies can be utilized to evaluate cellular immunogenicity, including:

1) Evaluation of primary T cell cultures from normal individuals (see, e.g., Wentworth, P. A. et al, Mol. Immunol. 32:603, 1995; Celis, E. et al., Proc. Natl. Acad. Sci. USA 91:2105, 1994; Tsai, V. et al., J. Immunol. 158:1796, 1997; Kawashima, I. et al., Human Immunol 59:1, 1998). This procedure involves the simulation of peripheral blood lymphocytes (PBL) from normal subjects with a test peptide in the presence of antigen presenting cells in vitro over a period of several weeks. T cells specific for the peptide become activated during this time and are detected using, e.g., a lymphokine- or ⁵¹Cr-release assay involving peptide sensitized target cells.

2) Immunization of HLA transgenic mice (see, e.g., Wentworth, P. A. et al., J. Immunol. 26:97, 1996; Wentworth, P. A. et al., Int. Immunol. 8:651, 1996; Alexander, J. et al., J. Immunol. 159:4753, 1997). For example, in such methods peptides in incomplete Freund's adjuvant are administered subcutaneously to HLA transgenic mice. Several weeks following immunization, splenocytes are removed and cultured in vitro in the presence of test peptide for approximately one week. Peptide-specific T cells are detected using, e.g., a ⁵¹Cr-release assay involving peptide sensitized target cells and target cells expressing endogenously generated antigen.

3) Demonstration of recall T cell responses from immune individuals who have been either effectively vaccinated and/or from chronically ill patients (see, e.g., Rehermann, B. et al., J. Exp. Med. 181:1047, 1995; Doolan, D. L. et al., Immunity 7:97, 1997; Bertoni, R. et al., J. Clin. Invest. 100:503, 1997; Threlkeld, S. C. et al., J. Immunol. 159:1648, 1997; Diepolder, H. M. et al., J. Virol. 71:6011, 1997). Accordingly, recall responses are detected by culturing PBL from subjects that have been exposed to the antigen due to disease and thus have generated an immune response “naturally”, or from patients who were vaccinated against the antigen. PBL from subjects are cultured in vitro for 1-2 weeks in the presence of test peptide plus antigen presenting cells (APC) to allow activation of “memory” T cells, as compared to “naive” T cells. At the end of the culture period, T cell activity is detected using assays including ⁵¹Cr release involving peptide-sensitized targets, T cell proliferation, or lymphokine release.

VI.) 98P4B6 TRANSGENIC ANIMALS

Nucleic acids that encode a 98P4B6-related protein can also be used to generate either transgenic animals or “knock out” animals that, in turn, are useful in the development and screening of therapeutically useful reagents. In accordance with established techniques, cDNA encoding 98P4B6 can be used to clone genomic. DNA that encodes 98P4B6. The cloned genomic sequences can then be used to generate transgenic animals containing cells that express DNA that encode 98P4B6. Methods for generating transgenic animals, particularly animals such as mice or rats, have become conventional in the art and are described, for example, in U.S. Pat. Nos. 4,736,866 issued 12 Apr. 1988, and 4,870,009 issued 26 Sep. 1989. Typically, particular cells would be targeted for 98P4B6 transgene incorporation with tissue-specific enhancers.

Transgenic animals that include a copy of a transgene encoding 98P4B6 can be used to examine the effect of increased expression of DNA that encodes 98P4B6. Such animals can be used as tester animals for reagents thought to confer protection from, for example, pathological conditions associated with its overexpression. In accordance with this aspect of the invention, an animal is treated with a reagent and a reduced incidence of a pathological condition, compared to untreated animals that bear the transgene, would indicate a potential therapeutic intervention for the pathological condition.

Alternatively, non-human homologues of 98P4B6 can be used to construct a 98P4B6 “knock out” animal that has a defective or altered gene encoding 98P4B6 as a result of homologous recombination between the endogenous gene encoding 98P4B6 and altered genomic DNA encoding 98P4B6 introduced into an embryonic cell of the animal. For example, cDNA that encodes 98P4B6 can be used to done genomic DNA encoding 98P4B6 in accordance with established techniques. A portion of the genomic DNA encoding 98P4B6 can be deleted or replaced with another gene, such as a gene encoding a selectable marker that can be used to monitor integration. Typically, several kilobases of unaltered flanking DNA (both at the 5′ and 3′ ends) are included in the vector (see, e.g., Thomas and Capecchi, Cell, 51:503 (1987) for a description of homologous recombination vectors). The vector is introduced into an embryonic stem cell line (e.g., by electroporation) and cells in which the introduced DNA has homologously recombined with the endogenous DNA are selected (see, e.g., Li et al., Cell, 69:915 (1992)). The selected cells are then injected into a blastocyst of an animal (e.g., a mouse or rat) to form aggregation chimeras (see, e.g., Bradley, in Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, E. J. Robertson, ed. (IRL, Oxford, 1987), pp. 113-152). A chimeric embryo can then be implanted into a suitable pseudopregnant female foster animal, and the embryo brought to term to create a “knock out” animal. Progeny harboring the homologously recombined DNA in their germ cells can be identified by standard techniques and used to breed animals in which all cells of the animal contain the homologously recombined DNA. Knock out animals can be characterized, for example, for their ability to defend against certain pathological conditions or for their development of pathological conditions due to absence of a 98P486 polypeptide.

VII.) METHODS FOR THE DETECTION Of 98P4B6

Another aspect of the present invention relates to methods for detecting 98P4B6 polynucleotides and 98P4B6-related proteins, as well as methods for identifying a cell that expresses 98P4B6. The expression profile of 98P4B6 makes it a diagnostic marker for metastasized disease. Accordingly, the status of 98P4B6 gene products provides information useful for predicting a variety of factors including susceptibility to advanced stage disease, rate of progression, and/or tumor aggressiveness. As discussed in detail herein, the status of 98P4B6 gene products in patent samples can be analyzed by a variety protocols that are well known in the art including immunohistochemical analysis, the variety of Northern blotting techniques including in situ hybridization, RT-PCR analysis (for example on laser capture micro-dissected samples), Western blot analysis and tissue array analysis.

More particularly, the invention provides assays for the detection of 98P4B6 polynucleotides in a biological sample, such as serum, bone, prostate, and other tissues, urine, semen, cell preparations, and the like. Detectable 98P4B6 polynucleotides include, for example, a 98P4B6 gene or fragment thereof, 98P4B6 mRNA, alternative splice variant 98P4B6 mRNAs, and recombinant DNA or RNA molecules that contain a 98P4B6 polynucleotide. A number of methods for amplifying and/or detecting the presence of 98P4B6 polynucleotides are well known in the art and can be employed in the practice of this aspect of the invention.

In one embodiment, a method for detecting a 98P4B6 mRNA in a biological sample comprises producing cDNA from the sample by reverse transcription using at least one primer; amplifying the cDNA so produced using a 98P4B6 polynucleotides as sense and antisense primers to amplify 98P486 cDNAs therein; and detecting the presence of the amplified 98P4B6 cDNA. Optionally, the sequence of the amplified 98P4B6 cDNA can be determined.

In another embodiment, a method of detecting a 98P4B6 gene in a biological sample comprises first isolating genomic DNA from the sample; amplifying the isolated genomic DNA using 98P4B6 polynucleotides as sense and antisense primers; and detecting the presence of the amplified 98P4B6 gene. Any number of appropriate sense and antisense probe combinations can be designed from a 98P4B6 nucleotide sequence (see, e.g., FIG. 2) and used for this purpose.

The invention also provides assays for detecting the presence of a 98P486 protein in a tissue or other biological sample such as serum, semen, bone, prostate, urine, cell preparations, and the like. Methods for detecting a 98P4B6-related protein are also well known and include, for example, immunoprecipitation, immunohistochemical analysis, Western blot analysis, molecular binding assays, ELISA, ELIFA and the like. For example, a method of detecting the presence of a 98P4B6-related protein in a biological sample comprises first contacting the sample with a 98P4B6 antibody, a 98P4B6-reactive fragment thereof, or a recombinant protein containing an antigen-binding region of a 98P4B6 antibody; and then detecting the binding of 98P4B6-related protein in the sample.

Methods for identifying a cell that expresses 98P4B6 are also within the scope of the invention. In one embodiment, an assay for identifying a cell that expresses a 98P4B6 gene comprises detecting the presence of 98P4B6 mRNA in the cell. Methods for the detection of particular mRNAs in cells are well known and include, for example, hybridization assays using complementary DNA probes (such as in situ hybridization using labeled 98P4B6 riboprobes, Northern blot and related techniques) and various nucleic acid amplification assays (such as RT-PCR using complementary primers specific for 98P4B6, and other amplification type detection methods, such as, for example, branched DNA, SISBA, TMA and the like). Alternatively, an assay for identifying a cell that expresses a 98P4B6 gene comprises detecting the presence of 98P4B6-related protein in the cell or secreted by the cell. Various methods for the detection of proteins are well known in the art and are employed for the detection of 98P4B6-related proteins and cells that express 98P4B6-related proteins.

98P4B6 expression analysis is also useful as a tool for identifying and evaluating agents that modulate 98P486 gene expression. For example, 98P4B6 expression is significantly upregulated in prostate cancer, and is expressed in cancers of the tissues listed in Table I. Identification of a molecule or biological agent that inhibits 98P4B6 expression or over-expression in cancer cells is of therapeutic value. For example, such an agent can be identified by using a screen that quantifies 98P4B6 expression by RT-PCR, nucleic acid hybridization or antibody binding.

VIII.) METHODS FOR MONITORING THE STATUS OF 98P4B6-RELATED GENES AND THEIR PRODUCTS

Oncogenesis is known to be a multistep process where cellular growth becomes progressively dysregulated and cells progress from a normal physiological state to precancerous and then cancerous states (see, e.g., Alers et al., Lab Invest. 77(5): 437-438 (1997) and Isaacs et al., Cancer Surv. 23: 19-32 (1995)). In this context, examining a biological sample for evidence of dysregulated cell growth (such as aberrant 98P4B6 expression in cancers) allows for early detection of such aberrant physiology, before a pathologic state such as cancer has progressed to a stage that therapeutic options are more limited and or the prognosis is worse. In such examinations, the status of 98P4B6 in a biological sample of interest can be compared, for example, to the status of 98P4B6 in a corresponding normal sample (e.g. a sample from that individual or alternatively another individual that is not affected by a pathology). An alteration in the status of 98P4B6 in the biological sample (as compared to the normal sample) provides evidence of dysregulated cellular growth. In addition to using a biological sample that is not affected by a pathology as a normal sample, one can also use a predetermined normative value such as a predetermined normal level of mRNA expression (see, e.g., Grever et al., J. Comp. Neurol. 1996 Dec. 9; 376(2): 306-14 and U.S. Pat. No. 5,837,501) to compare 98P4B6 status in a sample.

The term “status” in this context is used according to its art accepted meaning and refers to the condition or state of a gene and its products. Typically, skilled artisans use a number of parameters to evaluate the condition or state of a gene and its products. These include, but are not limited to the location of expressed gene products (including the location of 98P4B6 expressing cells) as well as the level, and biological activity of expressed gene products (such as 98P4B6 mRNA, polynucleotides and polypeptides). Typically, an alteration in the status of 98P4B6 comprises a change in the location of 98P4B6 and/or 98P4B6 expressing cells and/or an increase in 98P4B6 mRNA and/or protein expression.

98P4B6 status in a sample can be analyzed by a number of means well known in the art, including without limitation, immunohistochemical analysis, in situ hybridization, RT-PCR analysis on laser capture micro-dissected samples, Western blot analysis, and tissue array analysis. Typical protocols for evaluating the status of a 98P4B6 gene and gene products are found, for example in Ausubel et al. eds., 1995, Current Protocols In Molecular Biology, Units 2 (Northern Blotting), 4 (Southern Blotting), 15 (Immunoblotting) and 18 (PCR Analysis). Thus, the status of 98P4B6 in a biological sample is evaluated by various methods utilized by skilled artisans including, but not limited to genomic Southern analysis (to examine, for example perturbations in a 98P4B6 gene), Northern analysis and/or PCR analysis of 98P4B6 mRNA (to examine, for example alterations in the polynucleotide sequences or expression levels of 98P4B6 mRNAs), and, Western and/or immunohistochemical analysis (to examine, for example alterations in polypeptide sequences, alterations in polypeptide localization within a sample, alterations in expression levels of 98P4B6 proteins and/or associations of 98P4B6 proteins with polypeptide binding partners). Detectable 98P4B6 polynucleotides include, for example, a 98P4B6 gene or fragment thereof, 98P4B6 mRNA, alternative splice variants, 98P4B6 mRNAs, and recombinant DNA or RNA molecules containing a 98P4B6 polynucleotide.

The expression profile of 98P4B6 makes it a diagnostic marker for local and/or metastasized disease, and provides information on the growth or oncogenic potential of a biological sample. In particular, the status of 98P4B6 provides information useful for predicting-susceptibility to particular disease stages, progression, and/or tumor aggressiveness. The invention provides methods and assays for determining 98P4B6 status and diagnosing cancers that express 98P4B6, such as cancers of the tissues listed in Table I. For example, because 98P4B6 mRNA is so highly expressed in prostate and other cancers relative to normal prostate tissue, assays that evaluate the levels of 98P4B6 mRNA transcripts or proteins in a biological sample can be used to diagnose a disease associated with 98P4B6 dysregulation, and can provide prognostic information useful in defining appropriate therapeutic options.

The expression status of 98P4B6 provides information including the presence, stage and location of dysplastic, precancerous and cancerous cells, predicting susceptibility to various stages of disease, and/or for gauging tumor aggressiveness. Moreover, the expression profile makes it useful as an imaging reagent for metastasized disease. Consequently, an aspect of the invention is directed to the various molecular prognostic and diagnostic methods for examining the status of 98P4B6 in biological samples such as those from individuals suffering from, or suspected of suffering from a pathology characterized by dysregulated cellular growth, such as cancer.

As described above, the status of 98P4B6 in a biological sample can be examined by a number of well-known procedures in the art. For example, the status of 98P4B6 in a biological sample taken from a specific location in the body can be examined by evaluating the sample for the presence or absence of 98P4B6 expressing cells (e.g. those that express 98P4B6 mRNAs or proteins). This examination can provide evidence of dysregulated cellular growth, for example, when 98P4B6-expressing cells are found in a biological sample that does not normally contain such cells (such as a lymph node), because such alterations in the status of 98P4B6 in a biological sample are often associated with dysregulated cellular growth. Specifically, one indicator of dysregulated cellular growth is the metastases of cancer cells from an organ of origin (such as the prostate) to a different area of the body (such as a lymph node). In this context, evidence of dysregulated cellular growth is important for example because occult lymph node metastases can be detected in a substantial proportion of patients with prostate cancer, and such metastases are associated with known predictors of disease progression (see, e.g., Murphy et al., Prostate 42(4): 315-317 (2000); Su et al., Semin. Surg. Oncol. 18(1): 17-28 (2000) and Freeman et al., J Urol 1995 August 154(2 Pt 1):474-8).

In one aspect, the invention provides methods for monitoring 98P4B6 gene products by determining the status of 98P4B6 gene products expressed by cells from an individual suspected of having a disease associated with dysregulated cell growth (such as hyperplasia or cancer) and then comparing the status so determined to the status of 98P4B6 gene products in a corresponding normal sample. The presence of aberrant 98P4B6 gene products in the test sample relative to the normal sample provides an indication of the presence of dysregulated cell growth within the cells of the individual.

In another aspect, the invention provides assays useful in determining the presence of cancer in an individual, comprising detecting a significant increase in 98P4B6 mRNA or protein expression in a test cell or tissue sample relative to expression levels in the corresponding normal cell or tissue. The presence of 98P4B6 mRNA can, for example, be evaluated in tissues including but not limited to those listed in Table I. The presence of significant 98P4B6 expression in any of these tissues is useful to indicate the emergence, presence and/or severity of a cancer, since the corresponding normal tissues do not express 98P4B6 mRNA or express it at lower levels.

In a related embodiment, 98P4B6 status is determined at the protein level rather than at the nucleic acid level. For example, such a method comprises determining the level of 98P4B6 protein expressed by cells in a test issue sample and comparing the level so determined to the level of 98P4B6 expressed in a corresponding normal sample. In one embodiment, the presence of 98P4B6 protein is evaluated, for example, using immunohistochemical methods. 98P4B6 antibodies or binding partners capable of detecting 98P4B6 protein expression are used in a variety of assay formats well known in the art for this purpose.

In a further embodiment, one can evaluate the status of 98P4B6 nucleotide and amino acid sequences in a biological sample in order to identify perturbations in the structure of these molecules. These perturbations can include insertions, deletions, substitutions and the like. Such evaluations are useful because perturbations in the nucleotide and amino acid sequences are observed in a large number of proteins associated with a growth dysregulated phenotype (see, e.g., Marrogi et al., 1999, J. Cutan. Pathol. 26(8):369-378). For example, a mutation in the sequence of 98P4B6 may be indicative of the presence or promotion of a tumor. Such assays therefore have diagnostic and predictive value where a mutation in 98P4B6 indicates a potential loss of function or increase in tumor growth.

A wide variety of assays for observing perturbations in nucleotide and amino acid sequences are well known in the art. For example, the size and structure of nucleic acid or amino acid sequences of 98P4B6 gene products are observed by the Northern, Southern, Western, PCR and DNA sequencing protocols discussed herein. In addition, other methods for observing perturbations in nucleotide and amino acid sequences such as single strand conformation polymorphism analysis are well known in the art (see, e.g., U.S. Pat. Nos. 5,382,510 issued 7 Sep. 1999, and 5,952,170 issued 17 Jan. 1995).

Additionally, one can examine the methylation status of a 98P4B6 gene in a biological sample. Aberrant demethylation and/or hypermethylation of CpG islands in gene 5′ regulatory regions frequently occurs in immortalized and transformed cells, and can result in altered expression of various genes. For example, promoter hypermethylation of the pi-class glutathione S-transferase (a protein expressed in normal prostate but not expressed in >90% of prostate carcinomas) appears to permanently silence transcription of this gene and is the most frequently detected genomic alteration in prostate carcinomas (De Marzo et al., Am. J. Pathol. 155(6): 1985-1992 (1999)). In addition, this alteration is present in at least 70% of cases of high-grade prostatic intraepithelial neoplasia (PIN) (Brooks et al., Cancer Epidemiol. Biomarkers Prev., 1998, 7:531-536). In another example, expression of the LAGE-I tumor specific gene (which is not expressed in normal prostate but is expressed in 25-50% of prostate cancers) is induced by deoxy-azacytidine in lymphoblastoid cells, suggesting that tumoral expression is due to demethylation (Lethe et al., Int. J. Cancer 76(6): 903-908 (1998)). A variety of assays for examining methylation status of a gene are well known in the art. For example, one can utilize, in Southern hybridization approaches, methylation-sensitive restriction enzymes that cannot cleave sequences that contain methylated CpG sites to assess the methylation status of CpG islands. In addition, MSP (methylation specific PCR) can rapidly profile the methylation status of all the CpG sites present in a CpG island of a given gene. This procedure involves initial modification of DNA by sodium bisulfite (which will convert all unmethylated cytosines to uracil) followed by amplification using primers specific for methylated versus unmethylated DNA. Protocols involving methylation interference can also be found for example in Current Protocols In Molecular Biology, Unit 12, Frederick M. Ausubel et al. eds., 1995.

Gene amplification is an additional method for assessing the status of 98P4B6. Gene amplification is measured in a sample directly, for example, by conventional Southern blotting or Northern blotting to quantitate the transcription of mRNA (Thomas, 1980, Proc. Nail. Acad. Sci. USA, 77:5201-5205), dot blotting (DNA analysis), or in situ hybridization, using an appropriately labeled probe, based on the sequences provided herein. Alternatively, antibodies are employed that recognize specific duplexes, including DNA duplexes, RNA duplexes, and DNA-RNA hybrid duplexes or DNA-protein duplexes. The antibodies in turn are labeled and the assay carried out where the duplex is bound to a surface, so that upon the formation of duplex on the surface, the presence of antibody bound to the duplex can be detected.

Biopsied tissue or peripheral blood can be conveniently assayed for the presence of cancer cells using for example, Northern, dot blot or RT-PCR analysis to detect 98P4B6 expression. The presence of RT-PCR amplifiable 98P4B6 mRNA provides an indication of the presence of cancer. RT-PCR assays are well known in the art. RT-PCR detection assays for tumor cells in peripheral blood are currently being evaluated for use in the diagnosis and management of a number of human solid tumors. In the prostate cancer field, these include RT-PCR assays for the detection of cells expressing PSA and PSM (Verkaik et al., 1997, Urol. Res. 25:373-384; Ghossein et al., 1995, J. Clin. Oncol. 13:1195-2000; Heston et al., 1995, Clin. Chem. 41:1687-1688).

A further aspect of the invention is an assessment of the susceptibility that an individual has for developing cancer. In one embodiment, a method for predicting susceptibility to cancer comprises detecting 98P4B6 mRNA or 98P4B6 protein in a issue sample, its presence indicating susceptibility to cancer, wherein the degree of 98P4B6 mRNA expression correlates to the degree of susceptibility. In a specific embodiment, the presence of 98P4B6 in prostate or other tissue is examined, with the presence of 98P4B6 in the sample providing an indication of prostate cancer susceptibility (or the emergence or existence of a prostate tumor). Similarly, one can evaluate the integrity 98P4B6 nucleotide and amino acid sequences in a biological sample, in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like. The presence of one or more perturbations in 98P4B6 gene products in the sample is an indication of cancer susceptibility (or the emergence or existence of a tumor).

The invention also comprises methods for gauging tumor aggressiveness. In one embodiment, a method for gauging aggressiveness of a tumor comprises determining the level of 98P4B6 mRNA or 98P4B6 protein expressed by tumor cells, comparing the level so determined to the level of 98P4B6 mRNA or 98P4B6 protein expressed in a corresponding normal tissue taken from the same individual or a normal tissue reference sample, wherein the degree of 98P4B6 mRNA or 98P4B6 protein expression in the tumor sample relative to the normal sample indicates the degree of aggressiveness. In a specific embodiment, aggressiveness of a tumor is evaluated by determining the extent to which 98P4B6 is expressed in the tumor cells, with higher expression levels indicating more aggressive tumors. Another embodiment is the evaluation of the integrity of 98P4B6 nucleotide and amino acid sequences in a biological sample, in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like. The presence of one or more perturbations indicates more aggressive tumors.

Another embodiment of the invention is directed to methods for observing the progression of a malignancy in an individual over time. In one embodiment, methods for observing the progression of a malignancy in an individual over time comprise determining the level of 98P4B6 mRNA or 98P4B6 protein expressed by cells in a sample of the tumor, comparing the level so determined to the level of 98P4B6 mRNA or 98P4B6 protein expressed in an equivalent tissue sample taken from the same individual at a different time, wherein the degree of 98P4B6 mRNA or 98P4B6 protein expression in the tumor sample over time provides information on the progression of the cancer. In a specific embodiment, the progression of a cancer is evaluated by determining 98P4B6 expression in the tumor cells over time, where increased expression over time indicates a progression of the cancer. Also, one can evaluate the integrity 98P4B6 nucleotide and amino acid sequences in a biological sample in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like, where the presence of one or more perturbations indicates a progression of the cancer.

The above diagnostic approaches can be combined with any one of a wide variety of prognostic and diagnostic protocols known in the art. For example, another embodiment of the invention is directed to methods for observing a coincidence between the expression of 98P4B6 gene and 98P4B6 gene products (or perturbations in 98P4B6 gene and 98P4B6 gene products) and a factor that is associated with malignancy, as a means for diagnosing and prognosticating the status of a tissue sample. A wide variety of factors associated with malignancy can be utilized, such as the expression of genes associated with malignancy (e.g. PSA, PSCA and PSM expression for prostate cancer etc.) as well as gross cytological observations (see, e.g., Bocking et al., 1984, Anal. Quant. Cytol. 6(2):74-88; Epstein, 1995, Hum. Pathol. 26(2):223-9; Thorson et al., 1998, Mod. Pathol. 11 (6):543-51; Baisden et al., 1999, Am. J. Surg. Pathol. 23(8):918-24). Methods for observing a coincidence between the expression of 98P4B6 gene and 98P4B6 gene products (or perturbations in 98P4B6 gene and 98P4B6 gene products) and another factor that is associated with malignancy are useful, for example, because the presence of a set of specific factors that coincide with disease provides information crucial for diagnosing and prognosticating the status of a tissue sample.

In one embodiment, methods for observing a coincidence between the expression of 98P4B6 gene and 98P4B6 gene products (or perturbations in 98P4B6 gene and 98P4B6 gene products) and another factor associated with malignancy entails detecting the overexpression of 98P4B6 mRNA or protein in a issue sample, detecting the overexpression of PSA mRNA or protein in a tissue sample (or PSCA or PSM expression), and observing a coincidence of 98P4B6 mRNA or protein and PSA mRNA or protein overexpression (or PSCA or PSM expression). In a specific embodiment, the expression of 98P4B6 and PSA mRNA in prostate tissue is examined, where the coincidence of 98P4B6 and PSA mRNA overexpression in the sample indicates the existence of prostate cancer, prostate cancer susceptibility or the emergence or status of a prostate tumor.

Methods for detecting and quantifying the expression of 98P4B6 mRNA or protein are described herein, and standard nucleic acid and protein detection and quantification technologies are well known in the art Standard methods for the detection and quantification of 98P4B6 mRNA include in situ hybridization using labeled 98P4B6 riboprobes, Northern blot and related techniques using 98P4B6 polynucleotide probes, RT-PCR analysis using primers specific for 98P4B6, and other amplification type detection methods, such as, for example, branched DNA, SISBA, TMA and the like. In a specific embodiment, semi-quantitative RT-PCR is used to detect and quantify 98P4B6 mRNA expression. Any number of primers capable of amplifying 98P4B6 can be used for this purpose, including but not limited to the various primer sets specifically described herein. In a specific embodiment, polyclonal or monoclonal antibodies specifically reactive with the wild-type 98P486 protein can be used in an immunohistochemical assay of biopsied tissue.

IX.) IDENTIFICATION OF MOLECULES THAT INTERACT WITH 98P4B6

The 98P4B6 protein and nucleic acid sequences disclosed herein allow a skilled artisan to identify proteins, small molecules and other agents that interact with 98P4B6, as well as pathways activated by 98P486 via any one of a variety of art accepted protocols. For example, one can utilize one of the so-called interaction trap systems (also referred to as the “two-hybrid assay”). In such systems, molecules interact and reconstitute a transcription factor which directs expression of a reporter gene, whereupon the expression of the reporter gene is assayed. Other systems identify protein-protein interactions in vivo through reconstitution of a eukaryotic transcriptional activator, see, e.g., U.S. Pat. Nos. 5,955,280 issued 21 Sep. 1999, 5,925,523 issued 20 Jul. 1999, 5,846,722 issued 8 Dec. 1998 and 6,004,746 issued 21 Dec. 1999. Algorithms are also available in the art for genome-based predictions of protein function (see, e.g., Marcotte, et al., Nature 402: 4 Nov. 1999, 83-86).

Alternatively one can screen peptide libraries to identify molecules that interact with 98P4B6 protein sequences. In such methods, peptides that bind to 98P4B6 are identified by screening libraries that encode a random or controlled collection of amino acids. Peptides encoded by the libraries are expressed as fusion proteins of bacteriophage coat proteins, the bacteriophage particles are then screened against the 98P4B6 protein(s).

Accordingly, peptides having a wide variety of uses, such as therapeutic, prognostic or diagnostic reagents, are thus identified without any prior information on the structure of the expected ligand or receptor molecule. Typical peptide libraries and screening methods that can be used to identify molecules that interact with 98P4B6 protein sequences are disclosed for example in U.S. Pat. Nos. 5,723,286 issued 3 Mar. 1998 and 5,733,731 issued 31 Mar. 1998.

Alternatively, cell lines that express 98P4B6 are used to identify protein-protein interactions mediated by 98P4B6. Such interactions can be examined using immunoprecipitation techniques (see, e.g., Hamilton B. J., et al. Biochem. Biophys. Res. Commun. 1999, 261:646-51). 98P4B6 protein can be immunoprecipitated from 98P4B6-expressing cell lines using anti-98P4B6 antibodies. Alternatively, antibodies against His-tag can be used in a cell line engineered to express fusions of 98P4B6 and a His-tag (vectors mentioned above). The immunoprecipitated complex can be examined for protein association by procedures such as Western blotting, ³⁵S-methionine labeling of proteins, protein microsequencing, silver staining and two-dimensional gel electrophoresis.

Small molecules and ligands that interact with 98P4B6 can be identified through related embodiments of such screening assays. For example, small molecules can be identified that interfere with protein function, including molecules that interfere with 98P4B6's ability to mediate phosphorylation and de-phosphorylation, interaction with DNA or RNA molecules as an indication of regulation of cell cycles, second messenger signaling or tumorigenesis. Similarly, small molecules that modulate 98P4B6-related ion channel, protein pump, or cell communication functions are identified and used to treat patients that have a cancer that expresses 98P4B6 (see, e.g., Hille, B., Ionic Channels of Excitable Membranes 2^(nd) Ed., Sinauer Assoc., Sunderland, Mass., 1992). Moreover, ligands that regulate 98P4B6 function can be identified based on their ability to bind 98P4B6 and activate a reporter construct. Typical methods are discussed for example in U.S. Pat. No. 5,928,868 issued 27 Jul. 1999, and include methods for forming hybrid ligands in which at least one ligand is a small molecule. In an illustrative embodiment, cells engineered to express a fusion protein of 98P4B6 and a DNA-binding protein are used to co-express a fusion protein of a hybrid ligand/small molecule and a cDNA library transcriptional activator protein. The cells further contain a reporter gene, the expression of which is conditioned on the proximity of the first and second fusion proteins to each other, an event that occurs only if the hybrid ligand binds to target sites on both hybrid proteins. Those cells that express the reporter gene are selected and the unknown small molecule or the unknown ligand is identified. This method provides a means of identifying modulators, which activate or inhibit 98P4B6.

An embodiment of this invention comprises a method of screening for a molecule that interacts with a 98P4B6 amino acid sequence shown in FIG. 2 or FIG. 3, comprising the steps of contacting a population of molecules with a 98P4B6 amino acid sequence, allowing the population of molecules and the 98P4B6 amino acid sequence to interact under conditions that facilitate an interaction, determining the presence of a molecule that interacts with the 98P4B6 amino acid sequence, and then separating molecules that do not interact with the 98P4B6 amino acid sequence from molecules that do. In a specific embodiment, the method further comprises purifying, characterizing and identifying a molecule that interacts with the 98P4B6 amino acid sequence. The identified molecule can be used to modulate a function performed by 98P4B6. In a preferred embodiment, the 98P4B6 amino acid sequence is contacted with a library of peptides.

X.) THERAPEUTIC METHODS AND COMPOSITIONS

The identification of 98P486 as a protein that is normally expressed in a restricted set of tissues, but which is also expressed in prostate and other cancers, opens a number of therapeutic approaches to the treatment of such cancers. As contemplated herein, 98P4B6 functions as a transcription factor involved in activating tumor-promoting genes or repressing genes that block tumorigenesis.

Accordingly, therapeutic approaches that inhibit the activity of a 98P4B6 protein are useful for patients suffering from a cancer that expresses 98P4B6. These therapeutic approaches generally fall into two classes. One class comprises various methods for inhibiting the binding or association of a 98P4B6 protein with its binding partner or with other proteins. Another class comprises a variety of methods for inhibiting the transcription of a 98P4B6 gene or translation of 98P4B6 mRNA.

X.A.) Anti-Cancer Vaccines

The invention provides cancer vaccines comprising a 98P4B6-related protein or 98P4B6-related nucleic acid. In view of the expression of 98P4B6, cancer vaccines prevent and/or treat 98P4B6-expressing cancers with minimal or no effects on non-target tissues. The use of a tumor antigen in a vaccine that generates humoral and/or cell-mediated immune responses as ant cancer therapy is well known in the art and has been employed in prostate cancer using human PSMA and rodent PAP immunogens (Hodge et al., 1995, Int. J. Cancer 63:231-237; Fong et al., 1997, J. Immunol. 159:3113-3117).

Such methods can be readily practiced by employing a 98P4B6-related protein, or a 98P4B6-encoding nucleic acid molecule and recombinant vectors capable of expressing and presenting the 98P4B6 immunogen (which typically comprises a number of antibody or T cell epitopes). Skilled artisans understand that a wide variety of vaccine systems for delivery of immunoreactive epitopes are known in the art (see, e.g., Heryln et al., Ann Med 1999 Feb. 31(1):66-78; Maruyama et al., Cancer Immunol Immunother 2000 June 49(3):123-32) Briefly, such methods of generating an immune response (e.g. humoral and/or cell-mediated) in a mammal, comprise the steps of: exposing the mammal's immune system to an immunoreactive epitope (e.g. an epitope present in a 98P4B6 protein shown in FIG. 3 or analog or homolog thereof) so that the mammal generates an immune response that is specific for that epitope (e.g. generates antibodies that specifically recognize that epitope). In a preferred method, a 98P4B6 immunogen contains a biological motif, see e.g., Tables VIII-XXI and XXII-XLIX, or a peptide of a size range from 98P4B6 indicated in FIG. 5, FIG. 6, FIG. 7, FIG. 8, and FIG. 9.

The entire 98P4B6 protein, immunogenic regions or epitopes thereof can be combined and delivered by various means. Such vaccine compositions can include, for example, lipopeptides (e.g., Vibello, A. et al., J. Clin. Invest 95:341, 1995), peptide compositions encapsulated in poly(DL-lactide-co-glycolide) (“PLG”) microspheres (see, e.g., Eldridge, et al., Molec. Immunol. 28:287-294, 1991: Alonso et al., Vaccine 12:299-306, 1994; Jones et al., Vaccine 13:675-681, 1995), peptide compositions contained in immune stimulating complexes (ISCOMS) (see; e.g., Takahashi et al., Nature 344:873-875, 1990; Hu et al., Clin Exp Immunol. 113:235-243, 1998), multiple antigen peptide systems (MAPs) (see e.g., Tam, J. P., Proc. Natl. Acad. Sci. U.S.A. 85:5409-5413, 1988; Tam, J. P., J. Immunol. Methods 196:17-32, 1996), peptides formulated as multivalent peptides; peptides for use in ballistic delivery systems, typically crystallized peptides, viral delivery vectors (Perkus, M. E. et al., In: Concepts in vaccine development, Kaufmann, S. H. E., ed., p. 379, 1996; Chakrabarti, S. et al., Nature 320:535, 1986; Hu, S. L. et al., Nature 320:537, 1986; Kieny, M.-P. et al., AIDS Bio/Technology 4:790, 1986; Top, F. H. et al., J. Infect. Dis. 124:148, 1971; Chanda, P. K. et al., Virology 175:535, 1990), particles of viral or synthetic origin (e.g., Kofler, N. et al., J. Immunol. Methods. 192:25, 1996; Eldridge, J. H. et al., Sem. Hematol. 30:16, 1993; Falo, L. D., Jr. et al., Nature Med. 7:649, 1995), adjuvants (Warren, H. S., Vogel, F. R., and Chedid, L. A. Annu. Rev. Immunol. 4:369, 1986; Gupta, R. K. et al., Vaccine 11:293, 1993), liposomes (Reddy, R. et al., J. Immunol. 148:1585, 1992; Rock, K. L., Immunol. Today 17:131, 1996), or, naked or particle absorbed cDNA (Ulmer, J. B. et al., Science 259:1745, 1993; Robinson, H. L., Hunt, L. A., and Webster, R. G., Vaccine 11:957, 1993; Shiver, J. W. et al., In: Concepts in vaccine development, Kaufmann, S. H. E., ed., p. 423, 1996; Cease, K. B., and Berzofsky, J. A., Annu. Rev. Immunol. 12:923, 1994 and Eldridge, J. H. et al., Sem. Hematol. 30:16, 1993). Toxin-targeted delivery technologies, also known as receptor mediated targeting, such as those of Avant Immunotherapeutics, Inc. (Needham, Mass.) may also be used.

In patients with 98P4B6-associated cancer, the vaccine compositions of the invention can also be used in conjunction with other treatments used for cancer, e.g., surgery, chemotherapy, drug therapies, radiation therapies, etc. including use in combination with immune adjuvants such as IL-2, IL-12, GM-CSF, and the like.

Cellular Vaccines:

CTL epitopes can be determined using specific algorithms to identify peptides within 98P4B6 protein that bind corresponding HLA alleles (see e.g., Table IV; Epimer™ and Epimatrix™, Brown University; and, BIMAS. In a preferred embodiment, a 98P4B6 immunogen contains one or more amino acid sequences identified using techniques well known in the art, such as the sequences shown in Tables VIII-XXI and XXII-XLIX or a peptide of 8, 9, 10 or 11 amino acids specified by an HLA Class I motif/supermotif (e.g., Table IV (A), Table IV (D), or Table IV (E)) and/or a peptide of at least 9 amino adds that comprises an HLA Class II motif/supermotif (e.g., Table IV (B) or Table IV (C)). As is appreciated in the art, the HLA Class I binding groove is essentially closed ended so that peptides of only a particular size range can fit into the groove and be bound, generally HLA Class I epitopes are 8, 9, 10, or 11 amino acids long. In contrast, the HLA Class II binding groove is essentially open ended; therefore a peptide of about 9 or more amino acids can be bound by an HLA Class II molecule. Due to the binding groove differences between HLA Class I and II, HLA Class I motifs are length specific, i.e., position two of a Class I motif is the second amino acid in an amino to carboxyl direction of the peptide. The amino acid positions in a Class II motif are relative only to each other, not the overall peptide, i.e., additional amino acids can be attached to the amino and/or carboxyl termini of a motif-bearing sequence. HLA Class II epitopes are often 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids long, or longer than 25 amino acids.

Antibody-Based Vaccines

A wide variety of methods for generating an immune response in a mammal are known in the art (for example as the first step in the generation of hybridomas). Methods of generating an immune response in a mammal comprise exposing the mammal's immune system to an immunogenic epitope on a protein (e.g. a 98P4B6 protein) so that an immune response is generated. A typical embodiment consists of a method for generating an immune response to 98P4B6 in a host, by contacting the host with a sufficient amount of at least one 98P4B6 B cell or cytotoxic T-cell epitope or analog thereof; and at least one periodic interval thereafter re-contacting the host with the 98P4B6 B cell or cytotoxic T-cell epitope or analog thereof. A specific embodiment consists of a method of generating an immune response against a 98P4B6-related protein or a man-made multiepitopic peptide comprising: administering 98P4B6 immunogen (e.g. a 98P4B6 protein or a peptide fragment thereof, a 98P4B6 fusion protein or analog etc.) in a vaccine preparation to a human or another mammal. Typically, such vaccine preparations further contain a suitable adjuvant (see, e.g., U.S. Pat. No. 6,146,635) or a universal helper epitope such as a PADRE™ peptide (Epimmune Inc., San Diego, Calif.; see, e.g., Alexander et al., J. Immunol. 2000 164(3); 164(3): 1625-1633; Alexander et al., Immunity 1994 1(9): 751-761 and Alexander et al., Immunol. Res. 1998 18(2): 79-92). An alternative method comprises generating an immune response in an individual against a 98P4B6 immunogen by: administering in vivo to muscle or skin of the individual's body a DNA molecule that comprises a DNA sequence that encodes a 98P4B6 immunogen, the DNA sequence operatively linked to regulatory sequences which control the expression of the DNA sequence; wherein the DNA molecule is taken up by cells, the DNA sequence is expressed in the cells and an immune response is generated against the immunogen (see, e.g., U.S. Pat. No. 5,962,428). Optionally a genetic vaccine facilitator such as anionic lipids; saponins; lectins; estrogenic compounds; hydroxylated lower alkyls; dimethyl sulfoxide; and urea is also administered. In addition, an antiidiotypic antibody can be administered that mimics 98P4B6, in order to generate a response to the target antigen.

Nucleic Acid Vaccines:

Vaccine compositions of the invention include nucleic acid-mediated modalities. DNA or RNA that encode protein(s) of the invention can be administered to a patient. Genetic immunization methods can be employed to generate prophylactic or therapeutic humoral and cellular immune responses directed against cancer cells expressing 98P4B6. Constructs comprising DNA encoding a 98P4B6-related protein/immunogen and appropriate regulatory sequences can be injected directly into muscle or skin of an individual, such that the cells of the muscle or skin take-up the construct and express the encoded 98P486 protein/immunogen. Alternatively, a vaccine comprises a 98P4B6-related protein. Expression of the 98P4B6-related protein immunogen results in the generation of prophylactic or therapeutic humoral and cellular immunity against cells that bear a 98P4B6 protein. Various prophylactic and therapeutic genetic immunization techniques known in the art can be used (for review, see information and references published at Internet address genweb.com). Nucleic acid-based delivery is described, for instance, in Wolff et. al., Science 247:1465 (1990) as well as U.S. Pat. Nos. 5,580,859; 5,589,466; 5,804,566; 5,739,118; 5,736,524; 5,679,647; WO 98/04720. Examples of DNA-based delivery technologies include “naked DNA”, facilitated (bupivicaine, polymers, peptide-mediated) delivery, cationic lipid complexes, and particle-mediated (“gene gun”) or pressure-mediated delivery (see, e.g., U.S. Pat. No. 5,922,687).

For therapeutic or prophylactic immunization purposes, proteins of the invention can be expressed via viral or bacterial vectors. Various viral gene delivery systems that can be used in the practice of the invention include, but are not limited to, vaccinia, fowlpox, canarypox, adenovirus, influenza, poliovirus, adeno-associated virus, lentivirus, and sindbis virus (see, e.g., Restifo, 1996, Curr. Opin. Immunol. 8:658-663; Tsang et al. J. Natl. Cancer Inst. 87:982-990 (1995)). Non-viral delivery systems can also be employed by introducing naked DNA encoding a 98P4B6-related protein into the patient (e.g., intramuscularly or intradermally) to induce an ant-tumor response.

Vaccinia virus is used, for example, as a vector to express nucleotide sequences that encode the peptides of the invention. Upon introduction into a host, the recombinant vaccinia virus expresses the protein immunogenic peptide, and thereby elicits a host immune response. Vaccinia vectors and methods useful in immunization protocols are described in, e.g., U.S. Pat. No. 4,722,848. Another vector is BCG (Bacille Calmette Guerin). BCG vectors are described in Stover et al., Nature 351:456-460 (1991). A wide variety of other vectors useful for therapeutic administration or immunization of the peptides of the invention, e.g. adeno and adeno-associated virus vectors, retroviral vectors, Salmonella typhi vectors, detoxified anthrax toxin vectors, and the like, will be apparent to those skilled in the art from the description herein.

Thus, gene delivery systems are used to deliver a 98P4B6-related nucleic acid molecule. In one embodiment, the full-length human 98P4B6 cDNA is employed. In another embodiment, 98P4B6 nucleic acid molecules encoding specific cytotoxic T lymphocyte (CTL) and/or antibody epitopes are employed.

Ex Vivo Vaccines

Various ex vivo strategies can also be employed to generate an immune response. One approach involves the use of antigen presenting cells (APCs) such as dendritic cells (DC) to present 98P4B6 antigen to a patient's immune system. Dendritic cells express MHC class I and II molecules, B7 co-stimulator, and IL-12, and are thus highly specialized antigen presenting cells. In prostate cancer, autologous dendritic cells pulsed with peptides of the prostate-specific membrane antigen (PSMA) are being used in a Phase I clinical trial to stimulate prostate cancer patients' immune systems (Tjoa et al., 1996, Prostate 28:65-69; Murphy et al., 1996, Prostate 29:371-380). Thus, dendritic cells can be used to present 98P4B6 peptides to T cells in the context of MHC class I or II molecules. In one embodiment, autologous dendritic cells are pulsed with 98P4B6 peptides capable of binding to MHC class I and/or class II molecules. In another embodiment, dendritic cells are pulsed with the complete 98P4B6 protein. Yet another embodiment involves engineering the overexpression of a 98P4B6 gene in dendritic cells using various implementing vectors known in the art, such as adenovirus (Arthur et al., 1997, Cancer Gene Ther. 4:17-25), retrovirus (Henderson et al., 1996, Cancer Res. 56:3763-3770), lentivirus, adeno-associated virus, DNA transfection (Ribas et al., 1997, Cancer Res. 57:2865-2869), or tumor-derived RNA transfection (Ashley et al., 1997, J. Exp. Med. 186:1177-1182). Cells that express 98P4B6 can also be engineered to express immune modulators, such as GM-CSF, and used as immunizing agents.

X.B.) 98P4B6 as a Target for Antibody-Based Therapy

98P4B6 is an attractive target for antibody-based therapeutic strategies. A number of antibody strategies are known in the art for targeting both extracellular and intracellular molecules (see, e.g., complement and ADCC mediated killing as well as the use of intrabodies). Because 98P4B6 is expressed by cancer cells of various lineages relative to corresponding normal cells, systemic administration of 98P4B6-immunoreactive compositions are prepared that exhibit excellent sensitivity without toxic, non-specific and/or non-target effects caused by binding of the immunoreactive composition to non-target organs and tissues. Antibodies specifically reactive with domains of 98P4B6 are useful to treat 98P4B6-expressing cancers systemically, either as conjugates with a toxin or therapeutic agent, or as naked antibodies capable of inhibiting cell proliferation or function.

98P4B6 antibodies can be introduced into a patient such that the antibody binds to 98P4B6 and modulates a function, such as an interaction with a binding partner, and consequently mediates destruction of the tumor cells and/or inhibits the growth of the tumor cells. Mechanisms by which such antibodies exert a therapeutic effect can include complement-mediated cytolysis, antibody-dependent cellular cytotoxicity, modulation of the physiological function of 98P4B6, inhibition of ligand binding or signal transduction pathways, modulation of tumor cell differentiation, alteration of tumor angiogenesis factor profiles, and/or apoptosis.

Those skilled in the art understand that antibodies can be used to specifically target and bind immunogenic molecules such as an immunogenic region of a 98P4B6 sequence shown in FIG. 2 or FIG. 3. In addition, skilled artisans understand that it is routine to conjugate antibodies to cytotoxic agents (see, e.g., Slevers et al. Blood 93:11 3678-3684 (Jun. 1, 1999)). When cytotoxic and/or therapeutic agents are delivered directly to cells, such as by conjugating them to antibodies specific for a molecule expressed by that cell (e.g. 98P4B6), the cytotoxic agent will exert its known biological effect (i.e. cytotoxicity) on those cells.

A wide variety of compositions and methods for using antibody-cytotoxic agent conjugates to kill cells are known in the art. In the context of cancers, typical methods entail administering to an animal having a tumor a biologically effective amount of a conjugate comprising a selected cytotoxic and/or therapeutic agent linked to a targeting agent (e.g. an anti-98P4B6 antibody) that binds to a marker (e.g. 98P4B6) expressed, accessible to binding or localized on the cell surfaces. A typical embodiment is a method of delivering a cytotoxic and/or therapeutic agent to a cell expressing 98P4B6, comprising conjugating the cytotoxic agent to an antibody that immunospecifically binds to a 98P4B6 epitope, and, exposing the cell to the antibody-agent conjugate. Another illustrative embodiment is a method of treating an individual suspected of suffering from metastasized cancer, comprising a step of administering parenterally to said individual a pharmaceutical composition comprising a therapeutically effective amount of an antibody conjugated to a cytotoxic and/or therapeutic agent.

Cancer immunotherapy using anti-98P4B6 antibodies can be done in accordance with various approaches that have been successfully employed in the treatment of other types of cancer, including but not limited to colon cancer (Aden et al., 1998, Crit. Rev. Immunol. 18:133-138), multiple myeloma (Ozaki et al., 1997, Blood 90:3179-3186, Tsunenari et al., 1997, Blood 90:2437-2444), gastric cancer (Kasprzyk et al., 1992, Cancer Res. 52:2771-2776), B-cell lymphoma (Funakoshi et al., 1996, J. Immunother. Emphasis Tumor Immunol. 19:93-101), leukemia (Zhong et al., 1996, Leuk. Res. 20:581-589), colorectal cancer (Moun et al., 1994, Cancer Res. 54:6160-6166; Velders et al., 1995, Cancer Res. 55:4398-4403), and breast cancer (Shepard et al., 1991, J. Clin. Immunol. 11:117-127). Some therapeutic approaches involve conjugation of naked antibody to a toxin or radioisotope, such as the conjugation of ygi or 1131 to ant-CD20 antibodies (e.g., Zevalin™, IDEC Pharmaceuticals Corp. or Bexxar™, Coulter Pharmaceuticals), while others involve co-administration of antibodies and other therapeutic agents, such as Herceptin™ (trastuzumab) with paclitaxel (Genentech, Inc.). The antibodies can be conjugated to a therapeutic agent. To treat prostate cancer, for example, 98P4B6 antibodies can be administered in conjunction with radiation, chemotherapy or hormone ablation. Also, antibodies can be conjugated to a toxin such as calicheamicin (e.g., Mylotarg™, Wyeth-Ayerst, Madison, N.J., a recombinant humanized IgG₄ kappa antibody conjugated to antitumor antibiotic calicheamicin) or a maytansinoid (e.g., taxane-based Tumor-Activated Prodrug, TAP, platform, ImmunoGen, Cambridge, Mass., also see e.g., U.S. Pat. No. 5,416,064).

Although 98P4B6 antibody therapy is useful for all stages of cancer, antibody therapy can be particularly appropriate in advanced or metastatic cancers. Treatment with the antibody therapy of the invention is indicated for patients who have received one or more rounds of chemotherapy. Alternatively, antibody therapy of the invention is combined with a chemotherapeutic or radiation regimen for patients who have not received chemotherapeutic treatment. Additionally, antibody therapy can enable the use of reduced dosages of concomitant chemotherapy, particularly for patients who do not tolerate the toxicity of the chemotherapeutic agent very well. Fan et al. (Cancer Res. 53:4637-4642, 1993), Prewelt et al. (International J. of Onco. 9:217-224, 1996), and Hancock et al. (Cancer Res. 51:4575-4580, 1991) describe the use of various antibodies together with chemotherapeutic agents.

Although 98P4B6 antibody therapy is useful for all stages of cancer, antibody therapy can be particularly appropriate in advanced or metastatic cancers. Treatment with the antibody therapy of the invention is indicated for patients who have received one or more rounds of chemotherapy. Alternatively, antibody therapy of the invention is combined with a chemotherapeutic or radiation regimen for patients who have not received chemotherapeutic treatment. Additionally, antibody therapy can enable the use of reduced dosages of concomitant chemotherapy, particularly for patients who do not tolerate the toxicity of the chemotherapeutic agent very well.

Cancer patients can be evaluated for the presence and level of 98P4B6 expression, preferably using immunohistochemical assessments of tumor tissue, quantitative 98P4B6 imaging, or other techniques that reliably indicate the presence and degree of 98P4B6 expression. Immunohistochemical analysis of tumor biopsies or surgical specimens is preferred for this purpose. Methods for immunohistochemical analysis of tumor tissues are well known in the art.

Anti-98P4B6 monoclonal antibodies that treat prostate and other cancers include those that initiate a potent immune response against the tumor or those that are directly cytotoxic. In this regard, anti-98P4B6 monoclonal antibodies (mAbs) can elicit tumor cell lysis by either complement-mediated or antibody-dependent cell cytotoxicity (ADCC) mechanisms, both of which require an intact Fc portion of the immunoglobulin molecule for interaction with effector cell Fc receptor sites on complement proteins. In addition, anti-98P4B6 mAbs that exert a direct biological effect on tumor growth are useful to treat cancers that express 98P4B6. Mechanisms by which directly cytotoxic mAbs act include: inhibition of cell growth, modulation of cellular differentiation, modulation of tumor angiogenesis factor profiles, and the induction of apoptosis. The mechanism(s) by which a particular anti-98P4B6 mAb exerts an anti-tumor effect is evaluated using any number of in vitro assays that evaluate cell death such as ADCC, ADMMC, complement-mediated cell lysis, and so forth, as is generally known in the art.

In some patients, the use of murine or other non-human monoclonal antibodies, or human/mouse chimeric mAbs can induce moderate to strong immune responses against the non-human antibody. This can result in clearance of the antibody from circulation and reduced efficacy. In the most severe cases, such an immune response can lead to the extensive formation of immune complexes which, potentially, can cause renal failure. Accordingly, preferred monoclonal antibodies used in the therapeutic methods of the invention are those that are either fully human or humanized and that bind specifically to the target 98P4B6 antigen with high affinity but exhibit low or no antigenicity in the patient.

Therapeutic methods of the invention contemplate the administration of single anti-98P4B6 mAbs as well as combinations, or cocktails, of different mAbs. Such mAb cocktails can have certain advantages inasmuch as they contain mAbs that target different epitopes, exploit different effector mechanisms or combine directly cytotoxic mAbs with mAbs that rely on immune effector functionality. Such mAbs in combination can exhibit synergistic therapeutic effects. In addition, anti-98P4B6 mAbs can be administered concomitantly with other therapeutic modalities, including but not limited to various chemotherapeutic agents, androgen-blockers, immune modulators (e.g., IL-2, GM-CSF), surgery or radiation. The anti-98P4B6 mAbs are administered in their “naked” or unconjugated form, or can have a therapeutic agent(s) conjugated to them.

Ant-98P4B6 antibody formulations are administered via any route capable of delivering the antibodies to a tumor cell. Routes of administration include, but are not limited to, intravenous, intraperitoneal, intramuscular, intratumor, intradermal, and the like. Treatment generally involves repeated administration of the anti-98P4B6 antibody preparation, via an acceptable route of administration such as intravenous injection (IV), typically at a dose in the range of about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or 25 mg/kg body weight. In general, doses in the range of 10-1000 mg mAb per week are effective and well tolerated.

Based on clinical experience with the Herceptin™ mAb in the treatment of metastatic breast cancer, an initial loading dose of approximately 4 mg/kg patient body weight IV, followed by weekly doses of about 2 mg/kg IV of the anti-98P4B6 mAb preparation represents an acceptable dosing regimen. Preferably, the initial loading dose is administered as a 90-minute or longer infusion. The periodic maintenance dose is administered as a 30 minute or longer infusion, provided the initial dose was well tolerated. As appreciated by those of skill in the art, various factors can influence the ideal dose regimen in a particular case. Such factors include, for example, the binding affinity and half life of the Ab or mAbs used, the degree of 98P4B6 expression in the patient, the extent of circulating shed 98P4B6 antigen, the desired steady-state antibody concentration level, frequency of treatment, and the influence of chemotherapeutic or other agents used in combination with the treatment method of the invention, as well as the health status of a particular patient.

Optionally, patients should be evaluated for the levels of 98P4B6 in a given sample (e.g. the levels of circulating 98P4B6 antigen and/or 98P4B6 expressing cells) in order to assist in the determination of the most effective dosing regimen, etc. Such evaluations are also used for monitoring purposes throughout therapy, and are useful to gauge therapeutic success in combination with the evaluation of other parameters (for example, urine cytology and/or ImmunoCyt levels in bladder cancer therapy, or by analogy, serum PSA levels in prostate cancer therapy).

Anti-idiotypic anti-98P4B6 antibodies can also be used in anti-cancer therapy as a vaccine for inducing an immune response to cells expressing a 98P4B6-related protein. In particular, the generation of anti-idiotypic antibodies is well known in the art; this methodology can readily be adapted to generate anti-idiotypic anti-98P4B6 antibodies that mimic an epitope on a 98P4B6-related protein (see, for example, Wagner et al., 1997, Hybridoma 16: 33-40; Foon et al., 1995, J. Clin. Invest. 96:334-342; Hedyn et al., 1996, Cancer Immunol. Immunother. 43:65-76). Such an ant-idiotypic antibody can be used in cancer vaccine strategies.

X.C.) 98P4B6 as a Target for Cellular Immune Responses

Vaccines and methods of preparing vaccines that contain an immunogenically effective amount of one or more HLA-binding peptides as described herein are further embodiments of the invention. Furthermore, vaccines in accordance with the invention encompass compositions of one or more of the claimed peptides. A peptide can be present in a vaccine individually. Alternatively, the peptide can exist as a homopolymer comprising multiple copies of the same peptide, or as a heteropolymer of various peptides. Polymers have the advantage of increased immunological reaction and, where different peptide epitopes are used to make up the polymer, the additional ability to induce antibodies and/or CTLs that react with different antigenic determinants of the pathogenic organism or tumor-related peptide targeted for an immune response. The composition can be a naturally occurring region of an antigen or can be prepared, e.g., recombinantly or by chemical synthesis.

Carriers that can be used with vaccines of the invention are well known in the art, and include, e.g., thyroglobulin, albumins such as human serum albumin, tetanus toxoid, polyamino adds such as poly L-lysine, poly L-glutamic acid, influenza, hepatitis B virus core protein, and the like. The vaccines can contain a physiologically tolerable (i.e., acceptable) diluent such as water, or saline, preferably phosphate buffered saline. The vaccines also typically include an adjuvant Adjuvants such as incomplete Freund's adjuvant, aluminum phosphate, aluminum hydroxide, or alum are examples of materials well known in the art. Additionally, as disclosed herein, CTL responses can be primed by conjugating peptides of the invention to lipids, such as tipalmitoyl-5-glycerylcysteinlyseryl-serine (P₃CSS). Moreover, an adjuvant such as a synthetic cytosine-phosphorothiolated-guanine-containing (CpG) oligonucleotides has been found to increase CTL responses 10- to 100-fold. (see, e.g. Davila and Celis, J. Immunol. 165:539-547 (2000))

Upon immunization with a peptide composition in accordance with the invention, via injection, aerosol, oral, transdermal, transmucosal, intrapleural, intrathecal, or other suitable routes, the immune system of the host responds to the vaccine by producing large amounts of CTLs and/or HTLs specific for the desired antigen. Consequently, the host becomes at least partially immune to later development of cells that express or overexpress 98P4B6 antigen, or derives at least some therapeutic benefit when the antigen was tumor-associated.

In some embodiments, it may be desirable to combine the class I peptide components with components that induce or facilitate neutralizing antibody and or helper T cell responses directed to the target antigen. A preferred embodiment of such a composition comprises class I and class II epitopes in accordance with the invention. An alternative embodiment of such a composition comprises a class I and/or class II epitope in accordance with the invention, along with a cross reactive HTL epitope such as PADRE™ (Epimmune, San Diego, Calif.) molecule (described e.g., in U.S. Pat. No. 5,736,142).

A vaccine of the invention can also include antigen-presenting cells (APC), such as dendritic cells (DC), as a vehicle to present peptides of the invention. Vaccine compositions can be created in vitro, following dendritic cell mobilization and harvesting, whereby loading of dendritic cells occurs in vitro. For example, dendritic cells are transfected, e.g., with a minigene in accordance with the invention, or are pulsed with peptides. The dendritic cell can then be administered to a patient to elicit immune responses in vivo. Vaccine compositions, either DNA- or peptide-based, can also be administered in vivo in combination with dendritic cell mobilization whereby loading of dendritic cells occurs in vivo.

Preferably, the following principles are utilized when selecting an array of epitopes for inclusion in a polyepitopic composition for use in a vaccine, or for selecting discrete epitopes to be included in a vaccine and/or to be encoded by nucleic acids such as a minigene. It is preferred that each of the following principles be balanced in order to make the selection. The multiple epitopes to be incorporated in a given vaccine composition may be, but need not be, contiguous in sequence in the native antigen from which the epitopes are derived.

1.) Epitopes are selected which, upon administration, mimic immune responses that have been observed to be correlated with tumor clearance. For HLA Class I this includes 3-4 epitopes that come from at least one tumor associated antigen (TAA). For HLA Class II a similar rationale is employed; again 34 epitopes are selected from at least one TAA (see, e.g., Rosenberg et al., Science 278:1447-1450). Epitopes from one TAA may be used in combination with epitopes from one or more additional TAAs to produce a vaccine that targets tumors with varying expression patlems of frequently-expressed TAAs.

2.) Epitopes are selected that have the requisite binding affinity established to be correlated with immunogenicity: for HLA Class I an IC₅₀ of 500 nM or less, often 200 nM or less; and for Class II an IC₅₀ of 1000 nM or less.

3.) Sufficient supermotif bearing-peptides, or a sufficient array of allele-specific motif-bearing peptides, are selected to give broad population coverage. For example, it is preferable to have at least 80% population coverage. A Monte Carlo analysis, a statistical evaluation known in the art, can be employed to assess the breadth, or redundancy of, population coverage.

4.) When selecting epitopes from cancer-related antigens it is often useful to select analogs because the patient may have developed tolerance to the native epitope.

5.) Of particular relevance are epitopes referred to as “nested epitopes.” Nested epitopes occur where at least two epitopes overlap in a given peptide sequence. A nested peptide sequence can comprise B cell, HLA class I and/or HLA class II epitopes. When providing nested epitopes, a general objective is to provide the greatest number of epitopes per sequence. Thus, an aspect is to avoid providing a peptide that is any longer than the amino terminus of the amino terminal epitope and the carboxyl terminus of the carboxyl terminal epitope in the peptide. When providing a multi-epitopic sequence, such as a sequence comprising nested epitopes, it is generally important to screen the sequence in order to insure that it does not have pathological or other deleterious biological properties.

6.) If a polyepitopic protein is created, or when creating a minigene, an objective is to generate the smallest peptide that encompasses the epitopes of interest. This principle is similar, if not the same as that employed when selecting a peptide comprising nested epitopes. However, with an artificial polyepitopic peptide, the size minimization objective is balanced against the need to integrate any spacer sequences between epitopes in the polyepitopic protein. Spacer amino acid residues can, for example, be introduced to avoid junctional epitopes (an epitope recognized by the immune system, not present in the target antigen, and only created by the man-made juxtaposition of epitopes), or to facilitate cleavage between epitopes and thereby enhance epitope presentation. Junctional epitopes are generally to be avoided because the recipient may generate an immune response to that non-native epitope. Of particular concern is a junctional epitope that is a “dominant epitope.” A dominant epitope may lead to such a zealous response that immune responses to other epitopes are diminished or suppressed.

7.) Where the sequences of multiple variants of the same target protein are present, potential peptide epitopes can also be selected on the basis of their conservancy. For example, a criterion for conservancy may define that the entire sequence of an HLA class I binding peptide or the entire 9-mer core of a class II binding peptide be conserved in a designated percentage of the sequences evaluated for a specific protein antigen.

X.C.1. Minigene Vaccines

A number of different approaches are available which allow simultaneous delivery of multiple epitopes. Nucleic acids encoding the peptides of the invention are a particularly useful embodiment of the invention. Epitopes for inclusion in a minigene are preferably selected according to the guidelines set forth in the previous section. A preferred means of administering nucleic acids encoding the peptides of the invention uses minigene constructs encoding a peptide comprising one or multiple epitopes of the invention.

The use of multi-epitope minigenes is described below and in, Ishioka et al., J. Immunol. 162:3915-3925, 1999; An, L. and Whitton, J. L., J. Virol 71:2292, 1997; Thomson, S. A. et al., J. Immunol. 157:822, 1996; Whitton, J. L. et al., J. Virol. 67:348, 1993; Hanke, R. et al., Vaccine 16:426, 1998. For example, a multi-epitope DNA plasmid encoding supermotif- and/or motif-bearing epitopes derived 98P4B6, the PADRE® universal helper T cell epitope or multiple HTL epitopes from 98P4B6 (see e.g., Tables VIII-XXI and XXII to XLIX), and an endoplasmic reticulum-translocating signal sequence can be engineered. A vaccine may also comprise epitopes that are derived from other TAAs.

The immunogenicity of a multi-epitopic minigene can be confirmed in transgenic mice to evaluate the magnitude of CTL induction responses against the epitopes tested. Further, the immunogenicity of DNA-encoded epitopes in vivo can be correlated with the in vitro responses of specific CTL lines against target cells transfected with the DNA plasmid. Thus, these experiments can show that the minigene serves to both: 1.) generate a CTL response and 2.) that the induced CTLs recognized cells expressing the encoded epitopes.

For example, to create a DNA sequence encoding the selected epitopes (minigene) for expression in human cells, the amino acid sequences of the epitopes may be reverse translated. A human codon usage table can be used to guide the codon choice for each amino acid. These epitope-encoding DNA sequences may be directly adjoined, so that when translated, a continuous polypeptide sequence is created. To optimize expression and/or immunogenicity, additional elements can be incorporated into the minigene design. Examples of amino acid sequences that can be reverse translated and included in the minigene sequence include: HLA class I epitopes, HLA class II epitopes, antibody epitopes, a ubiquitination signal sequence, and/or an endoplasmic reticulum targeting signal. In addition, HLA presentation of CTL and HTL epitopes may be improved by including synthetic (e.g. poly-alanine) or naturally-occurring flanking sequences adjacent to the CTL or HTL epitopes; these larger peptides comprising the epitope(s) are within the scope of the invention.

The minigene sequence may be converted to DNA by assembling oligonucleotides that encode the plus and minus strands of the minigene. Overlapping oligonucleotides (30-100 bases long) may be synthesized, phosphorylated, purified and annealed under appropriate conditions using well known techniques. The ends of the oligonucleotides can be joined, for example, using T4 DNA ligase. This synthetic minigene, encoding the epitope polypeptide, can then be cloned into a desired expression vector.

Standard regulatory sequences well known to those of skill in the art are preferably included in the vector to ensure expression in the target cells. Several vector elements are desirable: a promoter with a down-stream cloning site for minigene insertion; a polyadenylation signal for efficient transcription termination; an E. coli origin of replication; and an E. coli selectable marker (e.g. ampicillin or kanamycin resistance). Numerous promoters can be used for this purpose, e.g., the human cytomegalovirus (hCMV) promoter. See, e.g., U.S. Pat. Nos. 5,580,859 and 5,589,466 for other suitable promoter sequences.

Additional vector modifications may be desired to optimize minigene expression and immunogenicity. In some cases, introns are required for efficient gene expression, and one or more synthetic or naturally-occurring introns could be incorporated into the transcribed region of the minigene. The inclusion of mRNA stabilization sequences and sequences for replication in mammalian cells may also be considered for increasing minigene expression.

Once an expression vector is selected, the minigene is cloned into the polylinker region downstream of the promoter. This plasmid is transformed into an appropriate E. coli strain, and DNA is prepared using standard techniques. The orientation and DNA sequence of the minigene, as well as all other elements included in the vector, are confirmed using restriction mapping and DNA sequence analysis. Bacterial cells harboring the correct plasmid can be stored as a master cell bank and a working cell bank.

In addition, immunostimulatory sequences (ISSs or CpGs) appear to play a role in the immunogenicity of DNA vaccines. These sequences may be included in the vector, outside the minigene coding sequence, if desired to enhance immunogenicity.

In some embodiments, a bi-cistronic expression vector which allows production of both the minigene-encoded epitopes and a second protein (included to enhance or decrease immunogenicity) can be used. Examples of proteins or polypeptides that could beneficially enhance the immune response if co-expressed include cytokines (e.g., IL-2, IL-12, GM-CSF), cytokine-inducing molecules (e.g., LeIF), costimulatory molecules, or for HTL responses, pan-DR binding proteins (PADRE™, Epimmune, San Diego, Calif.). Helper (HTL) epitopes can be joined to intracellular targeting signals and expressed separately from expressed CTL epitopes; this allows direction of the HTL epitopes to a cell compartment different than that of the CTL epitopes. If required, this could facilitate more efficient entry of HTL epitopes into the HLA class II pathway, thereby improving HTL induction. In contrast to HTL or CTL induction, specifically decreasing the immune response by co-expression of immunosuppressive molecules (e.g. TGF-β) may be beneficial in certain diseases.

Therapeutic quantities of plasmid DNA can be produced for example, by fermentation in E. coli, followed by purification. Aliquots from the working cell bank are used to inoculate growth medium, and grown to saturation in shaker flasks or a bioreactor according to well-known techniques. Plasmid DNA can be purified using standard bioseparation technologies such as solid phase anion-exchange resins supplied by QIAGEN, Inc. (Valencia, Calif.). If required, supercoiled DNA can be isolated from the open circular and linear forms using gel electrophoresis or other methods.

Purified plasmid DNA can be prepared for injection using a variety of formulations. The simplest of these is reconstitution of lyophilized DNA in sterile phosphate-buffer saline (PBS). This approach, known as “naked DNA,” is currently being used for intramuscular (IM) administration in clinical trials. To maximize the immunotherapeutic effects of minigene DNA vaccines, an alternative method for formulating purified plasmid DNA may be desirable. A variety of methods have been described, and new techniques may become available. Cationic lipids, glycolipids, and fusogenic liposomes can also be used in the formulation (see, e.g., as described by WO 93/24640; Mannino & Gould-Fogerite, BioTechniques 6(7): 682 (1988); U.S. Pat. No. 5,279,833; WO 91/06309; and Felgner, et al., Proc. Nat'l Acad. Sci. USA 84:7413 (1987). In addition, peptides and compounds referred to collectively as protective, interactive, non-condensing compounds (PINC) could also be complexed to purified plasmid DNA to influence variables such as stability, intramuscular dispersion, or trafficking to specific organs or cell types.

Target cell sensitization can be used as a functional assay for expression and HLA class I presentation of minigene-encoded CTL epitopes. For example, the plasmid DNA is introduced into a mammalian cell line that is suitable as a target for standard CTL chromium release assays. The transfection method used will be dependent on the final formulation. Electroporation can be used for “naked” DNA, whereas cationic lipids allow direct in vitro transfection. A plasmid expressing green fluorescent protein (GFP) can be co-transfected to allow enrichment of transfected cells using fluorescence activated cell sorting (FACS). These cells are then chromium-51 (⁵¹Cr) labeled and used as target cells for epitope-specific CTL lines; cytolysis, detected by ⁵¹Cr release, indicates both production of, and HLA presentation of, minigene-encoded CTL epitopes. Expression of HTL epitopes may be evaluated in an analogous manner using assays to assess HTL activity.

In vivo immunogenicity is a second approach for functional testing of minigene DNA formulations. Transgenic mice expressing appropriate human HLA proteins are immunized with the DNA product. The dose and route of administration are formulation dependent (e.g., IM for DNA in PBS, intraperitoneal (i.p.) for lipid-complexed DNA). Twenty-one days after immunization, splenocytes are harvested and restimulated for one week in the presence of peptides encoding each epitope being tested. Thereafter, for CTL effector cells, assays are conducted for cytolysis of peptide-loaded, ⁵¹Cr-labeled target cells using standard techniques. Lysis of target cells that were sensitized by HLA loaded with peptide epitopes, corresponding to minigene-encoded epitopes, demonstrates DNA vaccine function for in vivo induction of CTLs. Immunogenicity of HTL epitopes is confirmed in transgenic mice in an analogous manner.

Alternatively, the nucleic acids can be administered using ballistic delivery as described, for instance, in U.S. Pat. No. 5,204,253. Using this technique, particles comprised solely of DNA are administered. In a further alternative embodiment, DNA can be adhered to particles, such as gold particles.

Minigenes can also be delivered using other bacterial or viral delivery systems well known in the art, e.g., an expression construct encoding epitopes of the invention can be incorporated into a viral vector such as vaccinia.

X.C.2. Combinations of CTL Peptides with Helper Peptides

Vaccine compositions comprising CTL peptides of the invention can be modified, e.g., analoged, to provide desired attributes, such as improved serum half life, broadened population coverage or enhanced immunogenicity.

For instance, the ability of a peptide to induce CTL activity can be enhanced by linking the peptide to a sequence which contains at least one epitope that is capable of inducing a T helper cell response. Although a CTL peptide can be directly linked to a T helper peptide, often CTL epitope/HTL epitope conjugates are linked by a spacer molecule. The spacer is typically comprised of relatively small, neutral molecules, such as amino adds or amino acid mimetics, which are substantially uncharged under physiological conditions. The spacers are typically selected from, e.g., Ala, Gly, or other neutral spacers of nonpolar amino adds or neutral polar amino adds. It will be understood that the optionally present spacer need not be comprised of the same residues and thus may be a hetero- or homo-oligomer. When present, the spacer will usually be at least one or two residues, more usually three to six residues and sometimes 10 or more residues. The CTL peptide epitope can be linked to the T helper peptide epitope either directly or via a spacer either at the amino or carboxy terminus of the CTL peptide. The amino terminus of either the immunogenic peptide or the T helper peptide may be acylated.

In certain embodiments, the T helper peptide is one that is recognized by T helper cells present in a majority of a genetically diverse population. This can be accomplished by selecting peptides that bind to many, most, or all of the HLA class II molecules. Examples of such amino acid bind many HLA Class II molecules include sequences from antigens such as tetanus toxoid at positions 830-843 (QYIKANSKFIGITE; SEQ ID NO: 97), Plasmodium falciparum circumsporozoite (CS) protein at positions 378-398 (DIEKKIAKMEKASSVFNWNS; SEQ ID NO: 98), and Streptococcus 18 kD protein at positions 116-131 (GAVDSILGGVATYGAA; SEQ ID NO: 99). Other examples include peptides bearing a DR 1-4-7 supermotif, or either of the DR3 motifs.

Alternatively, it is possible to prepare synthetic peptides capable of stimulating T helper lymphocytes, in a loosely HLA-restricted fashion, using amino acid sequences not found in nature (see, e.g., PCT publication WO 95/07707). These synthetic compounds called Pan-DR-binding epitopes (e.g., PADRE™, Epimmune, Inc., San Diego, Calif.) are designed, most preferably, to bind most HLA-DR (human HLA class II) molecules. For instance, a pan-DR-binding epitope peptide having the formula: XKXVAAVTLKAAX (SEQ ID NO: 100), where “X” is either cyclohexylalanine, phenylalanine, or tyrosine, and a is either D-alanine or L-alanine, has been found to bind to most HLA-DR alleles, and to stimulate the response of T helper lymphocytes from most individuals, regardless of their HLA type. An alternative of a pan-DR binding epitope comprises all “L” natural amino acids and can be provided in the form of nucleic acids that encode the epitope.

HTL peptide epitopes can also be modified to alter their biological properties. For example, they can be modified to include D-amino acids to increase their resistance to proteases and thus extend their serum half life, or they can be conjugated to other molecules such as lipids, proteins, carbohydrates, and the like to increase their biological activity. For example, a T helper peptide can be conjugated to one or more palmitic acid chains at either the amino or carboxyl termini.

X.C.3. Combinations of CTL Peptides with T Cell Priming Agents

In some embodiments it may be desirable to include in the pharmaceutical compositions of the invention at least one component which primes B lymphocytes or T lymphocytes. Lipids have been identified as agents capable of priming CTL in vivo. For example, palmitic acid residues can be attached to the ε- and α-amino groups of a lysine residue and then linked, e.g., via one or more linking residues such as Gly, Gly-Gly-, Ser, Ser-Ser, or the like, to an immunogenic peptide. The lipidated peptide can then be administered either directly in a micelle or particle, incorporated into a liposome, or emulsified in an adjuvant, e.g., incomplete Freund's adjuvant. In a preferred embodiment, a particularly effective immunogenic composition comprises palmitic acid attached to ε- and α-amino groups of Lys, which is attached via linkage, e.g., Ser-Ser, to the amino terminus of the immunogenic peptide.

As another example of lipid priming of CTL responses, E. coli lipoproteins, such as tripalmitoyl-S-glycerylcysteinlyseryl-serine (P₃CSS) can be used to prime virus specific CTL when covalently attached to an appropriate peptide (see, e.g., Deres, et al., Nature 342:561, 1989). Peptides of the invention can be coupled to P₃CSS, for example, and the lipopeptide administered to an individual to prime specifically an immune response to the target antigen. Moreover, because the induction of neutralizing antibodies can also be primed with P₃CSS-conjugated epitopes, two such compositions can be combined to more effectively elicit both humoral and cell-mediated responses.

X.C.4. Vaccine Compositions Comprising DC Pulsed with CTL and/or HTL Peptides

An embodiment of a vaccine composition in accordance with the invention comprises ex vivo administration of a cocktail of epitope-bearing peptides to PBMC, or isolated DC therefrom, from the pabent's blood. A pharmaceutical to facilitate harvesting of DC can be used, such as Progenipoietin™ (Pharmacia-Monsanto, St. Louis, Mo.) or GM-CSF/IL-4. After pulsing the DC with peptides and prior to reinfusion into patients, the DC are washed to remove unbound peptides. In this embodiment, a vaccine comprises peptide-pulsed DCs which present the pulsed peptide epitopes complexed with HLA molecules on their surfaces.

The DC can be pulsed ex vivo with a cocktail of peptides, some of which stimulate CTL responses to 98P4B6. Optionally, a helper T cell (HTL) peptide, such as a natural or artificial loosely restricted HLA Class II peptide, can be included to facilitate the CTL response. Thus, a vaccine in accordance with the invention is used to treat a cancer which expresses or overexpresses 98P4B6.

X.D. Adoptive Immunotherapy

Antigenic 98P4B6-related peptides are used to elicit a CTL and/or HTL response ex vivo, as well. The resulting CTL or HTL cells, can be used to treat tumors in patients that do not respond to other conventional forms of therapy, or will not respond to a therapeutic vaccine peptide or nucleic acid in accordance with the invention. Ex vivo CTL or HTL responses to a particular antigen are induced by incubating in tissue culture the patient's, or genetically compatible, CTL or HTL precursor cells together with a source of antigen-presenting cells (APC), such as dendritic cells, and the appropriate immunogenic peptide. After an appropriate incubation time (typically about 7-28 days), in which the precursor cells are activated and expanded into effector cells, the cells are infused back into the patient, where they will destroy (CTL) or facilitate destruction (HTL) of their specific target cell (e.g., a tumor cell). Transfected dendritic cells may also be used as antigen presenting cells.

X.E. Administration of Vaccines for Therapeutic or Prophylactic Purposes

Pharmaceutical and vaccine compositions of the invention are typically used to treat and/or prevent a cancer that expresses or overexpresses 98P4B6. In therapeutic applications, peptide and/or nucleic acid compositions are administered to a patient in an amount sufficient to elicit an effective B cell, CTL and/or HTL response to the antigen and to cure or at least partially arrest or slow symptoms and/or complications. An amount adequate to accomplish this is defined as “therapeutically effective dose.” Amounts effective for this use will depend on, e.g., the particular composition administered, the manner of administration, the stage and severity of the disease being treated, the weight and general state of health of the patient, and the judgment of the prescribing physician.

For pharmaceutical compositions, the immunogenic peptides of the invention, or DNA encoding them, are generally administered to an individual already bearing a tumor that expresses 98P4B6. The peptides or DNA encoding them can be administered individually or as fusions of one or more peptide sequences. Patients can be treated with the immunogenic peptides separately or in conjunction with other treatments, such as surgery, as appropriate.

For therapeutic use, administration should generally begin at the first diagnosis of 98P4B6-associated cancer. This is followed by boosting doses until at least symptoms are substantially abated and for a period thereafter. The embodiment of the vaccine composition (i.e., including, but not limited to embodiments such as peptide cocktails, polyepitopic polypeptides, minigenes, or TAA-specific CTLs or pulsed dendritic cells) delivered to the patient may vary according to the stage of the disease or the patient's health status. For example, in a patient with a tumor that expresses 98P4B6, a vaccine comprising 98P4B6-specific CTL may be more efficacious in killing tumor cells in patient with advanced disease than alternative embodiments.

It is generally important to provide an amount of the peptide epitope delivered by a mode of administration sufficient to stimulate effectively a cytotoxic T cell response; compositions which stimulate helper T cell responses can also be given in accordance with this embodiment of the invention.

The dosage for an initial therapeutic immunization generally occurs in a unit dosage range where the lower value is about 1, 5, 50, 500, or 1,000 μg and the higher value is about 10,000; 20,000; 30,000; or 50,000 μg. Dosage values for a human typically range from about 500 μg to about 50,000 μg per 70 kilogram patient. Boosting dosages of between about 1.0 μg to about 50,000 μg of peptide pursuant to a boosting regimen over weeks to months may be administered depending upon the patients response and condition as determined by measuring the specific activity of CTL and HTL obtained from the patient's blood. Administration should continue until at least clinical symptoms or laboratory tests indicate that the neoplasia, has been eliminated or reduced and for a period thereafter. The dosages, routes of administration, and dose schedules are adjusted in accordance with methodologies known in the art.

In certain embodiments, the peptides and compositions of the present invention are employed in serious disease states, that is, life-threatening or potentially life threatening situations. In such cases, as a result of the minimal amounts of extraneous substances and the relative nontoxic nature of the peptides in preferred compositions of the invention, it is possible and may be felt desirable by the treating physician to administer substantial excesses of these peptide compositions relative to these stated dosage amounts.

The vaccine compositions of the invention can also be used purely as prophylactic agents. Generally the dosage for an initial prophylactic immunization generally occurs in a unit dosage range where the lower value is about 1, 5, 50, 500, or 1000 μg and the higher value is about 10,000; 20,000; 30,000; or 50,000 μg. Dosage values for a human typically range from about 500 μg to about 50,000 μg per 70 kilogram patient. This is followed by boosting dosages of between about 1.0 μg to about 50,000 μg of peptide administered at defined intervals from about four weeks to six months after the initial administration of vaccine. The immunogenicity of the vaccine can be assessed by measuring the specific activity of CTL and HTL obtained from a sample of the patient's blood.

The pharmaceutical compositions for therapeutic treatment are intended for parenteral, topical, oral, nasal, intrathecal, or local (e.g. as a cream or topical ointment) administration. Preferably, the pharmaceutical compositions are administered parentally, e.g., intravenously, subcutaneously, intradermally, or intramuscularly. Thus, the invention provides compositions for parenteral administration which comprise a solution of the immunogenic peptides dissolved or suspended in an acceptable carrier, preferably an aqueous carrier.

A variety of aqueous carriers may be used, e.g., water, buffered water, 0.8% saline, 0.3% glycine, hyaluronic acid and the like. These compositions may be sterilized by conventional, well-known sterilization techniques, or may be sterile filtered. The resulting aqueous solutions may be packaged for use as is, or lyophilized, the lyophilized preparation being combined with a sterile solution prior to administration.

The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH-adjusting and buffering agents, tonicity adjusting agents, wetting agents, preservatives, and the like, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.

The concentration of peptides of the invention in the pharmaceutical formulations can vary widely, i.e., from less than about 0.1%, usually at or at least about 2% to as much as 20% to 50% or more by weight, and will be selected primarily by fluid volumes, viscosities, etc., in accordance with the particular mode of administration selected.

A human unit dose form of a composition is typically included in a pharmaceutical composition that comprises a human unit dose of an acceptable carrier, in one embodiment an aqueous carrier, and is administered in a volume/quantity that is known by those of skill in the art to be used for administration of such compositions to humans (see, e.g., Remington's Pharmaceutical Sciences, 17^(th) Edition, A. Gennaro, Editor, Mack Publishing Co., Easton, Pa., 1985). For example a peptide dose for initial immunization can be from about 1 to about 50,000 μg, generally 100-5,000 μg, for a 70 kg patient For example, for nucleic acids an initial immunization may be performed using an expression vector in the form of naked nucleic acid administered IM (or SC or ID) in the amounts of 0.5-5 mg at multiple sites. The nucleic acid (0.1 to 1000 μg) can also be administered using a gene gun. Following an incubation period of 3-4 weeks, a booster dose is then administered. The booster can be recombinant fowlpox virus administered at a dose of 5-10⁷ to 5×10⁹ pfu.

For antibodies, a treatment generally involves repeated administration of the anti-98P4B6 antibody preparation, via an acceptable route of administration such as intravenous injection (IV), typically at a dose in the range of about 0.1 to about 10 mg/kg body weight. In general, doses in the range of 10-500 mg mAb per week are effective and well tolerated. Moreover, an initial loading dose of approximately 4 mg/kg patient body weight IV, followed by weekly doses of about 2 mg/kg IV of the anti-98P4B6 mAb preparation represents an acceptable dosing regimen. As appreciated by those of skill in the art, various factors can influence the ideal dose in a particular case. Such factors include, for example, half life of a composition, the binding affinity of an Ab, the immunogenicity of a substance, the degree of 98P4B6 expression in the patient, the extent of circulating shed 98P4B6 antigen, the desired steady-state concentration level, frequency of treatment, and the influence of chemotherapeutic or other agents used in combination with the treatment method of the invention, as well as the health status of a particular patient. Non-limiting preferred human unit doses are, for example, 500 μg-1 mg, 1 mg-50 mg, 50 mg-100 mg, 100 mg-200 mg, 200 mg-300 mg, 400 mg-500 mg, 500 mg-600 mg, 600 mg-700 mg, 700 mg-800 mg, 800 mg-900 mg, 900 mg-1 g, or 1 mg-700 mg. In certain embodiments, the dose is in a range of 2-5 mg/kg body weight, e.g., with follow on weekly doses of 1-3 mg/kg; 0.5 mg, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 mg/kg body weight followed, e.g., in two, three or four weeks by weekly doses; 0.5-10 mg/kg body weight, e.g., followed in two, three or four weeks by weekly doses; 225, 250, 275, 300, 325, 350, 375, 400 mg m² of body area weekly; 1-600 mg m² of body area weekly, 225-400 mg m² of body area weekly; these does can be followed by weekly doses for 2, 3, 4, 5, 6, 7, 8, 9, 19, 11, 12 or more weeks.

In one embodiment, human unit dose forms of polynucleotides comprise a suitable dosage range or effective amount that provides any therapeutic effect. As appreciated by one of ordinary skill in the art a therapeutic effect depends on a number of factors, including the sequence of the polynucleotide, molecular weight of the polynucleotide and route of administration. Dosages are generally selected by the physician or other health care professional in accordance with a variety of parameters known in the art, such as severity of symptoms, history of the patient and the like. Generally, for a polynucleotide of about 20 bases, a dosage range may be selected from, for example, an independently selected lower limit such as about 0.1, 0.25, 0.5, 1, 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400 or 500 mg/kg up to an independently selected upper limit, greater than the lower limit, of about 60, 80, 100, 200, 300, 400, 500, 750, 1000, 1500, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000 or 10,000 mg/kg. For example, a dose may be about any of the following: 0.1 to 100 mg/kg, 0.1 to 50 mg/kg, 0.1 to 25 mg/kg, 0.1 to 10 mg/kg, 1 to 500 mg/kg, 100 to 400 mg/kg, 200 to 300 mg/kg, 1 to 100 mg/kg, 100 to 200 mg/kg, 300 to 400 mg/kg, 400 to 500 mg/kg, 500 to 1000 mg/kg, 500 to 5000 mg/kg, or 500 to 10,000 mg/kg. Generally, parenteral routes of administration may require higher doses of polynucleotide compared to more direct application to the nucleotide to diseased tissue, as do polynucleotides of increasing length.

In one embodiment, human unit dose forms of T-cells comprise a suitable dosage range or effective amount that provides any therapeutic effect. As appreciated by one of ordinary skill in the art, a therapeutic effect depends on a number of factors. Dosages are generally selected by the physician or other health care professional in accordance with a variety of parameters known in the art, such as severity of symptoms, history of the patient and the like. A dose may be about 10⁴ cells to about 10⁶ cells, about 10⁶ cells to about 10⁸ cells, about 10⁸ to about 10¹¹ cells, or about 10⁸ to about 5×10¹⁰ cells. A dose may also about 10⁶ cells/m² to about 10⁸ cells/m², or about 10⁶ cells/m² to about 10⁸ cells/m².

Proteins(s) of the invention, and/or nucleic acids encoding the protein(s), can also be administered via liposomes, which may also serve to: 1) target the proteins(s) to a particular tissue, such as lymphoid tissue; 2) to target selectively to diseases cells; or, 3) to increase the half-life of the peptide composition. Liposomes include emulsions, foams, micelles, insoluble monolayers, liquid crystals, phospholipid dispersions, lamellar layers and the like. In these preparations, the peptide to be delivered is incorporated as part of a liposome, alone or in conjunction with a molecule which binds to a receptor prevalent among lymphoid cells, such as monoclonal antibodies which bind to the CD45 antigen, or with other therapeutic or immunogenic compositions. Thus, liposomes either filled or decorated with a desired peptide of the invention can be directed to the site of lymphoid cells, where the liposomes then deliver the peptide compositions. Liposomes for use in accordance with the invention are formed from standard vesicle-forming lipids, which generally include neutral and negatively charged phospholipids and a sterol, such as cholesterol. The selection of lipids is generally guided by consideration of, e.g., liposome size, add lability and stability of the liposomes in the blood stream. A variety of methods are available for preparing liposomes, as described in, e.g., Szoka, et al., Ann. Rev. Biophys. Bioeng. 9:467 (1980), and U.S. Pat. Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

For targeting cells of the immune system, a ligand to be incorporated into the liposome can include, e.g., antibodies or fragments thereof specific for cell surface determinants of the desired immune system cells. A liposome suspension containing a peptide may be administered intravenously, locally, topically, etc. in a dose which varies according to, inter alia, the manner of administration, the peptide being delivered, and the stage of the disease being treated.

For solid compositions, conventional nontoxic solid carriers may be used which include, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, magnesium carbonate, and the like. For oral administration, a pharmaceutically acceptable nontoxic composition is formed by incorporating any of the normally employed excipients, such as those carriers previously listed, and generally 10-95% of active ingredient, that is, one or more peptides of the invention, and more preferably at a concentration of 25%-75%.

For aerosol administration, immunogenic peptides are preferably supplied in finely divided form along with a surfactant and propellant. Typical percentages of peptides are about 0.01%-20% by weight, preferably about 1%-10%. The surfactant must, of course, be nontoxic, and preferably soluble in the propellant. Representative of such agents are the esters or partial esters of fatty acids containing from about 6 to 22 carbon atoms, such as caproic, octanoic, lauric, palmitic, stearic, linoleic, linolenic, olesteric and oleic acids with an aliphatic polyhydric alcohol or its cyclic anhydride. Mixed esters, such as mixed or natural glycerides may be employed. The surfactant may constitute about 0.1%-20% by weight of the composition, preferably about 0.25-5%. The balance of the composition is ordinarily propellant. A carrier can also be included, as desired, as with, e.g., lecithin for intranasal delivery.

XI.) DIAGNOSTIC AND PROGNOSTIC EMBODIMENTS OF 98P4B6.

As disclosed herein, 98P4B6 polynucleotides, polypeptides, reactive cytotoxic T cells (CTL), reactive helper T cells (HTL) and anti-polypeptide antibodies are used in well known diagnostic, prognostic and therapeutic assays that examine conditions associated with dysregulated cell growth such as cancer, in particular the cancers listed in Table I (see, e.g., both its specific pattern of tissue expression as well as its overexpression in certain cancers as described for example in the Example entitled “Expression analysis of 98P4B6 in normal tissues, and patient specimens”).

98P4B6 can be analogized to a prostate associated antigen PSA, the archetypal marker that has been used by medical practitioners for years to identify and monitor the presence of prostate cancer (see, e.g., Merrill et al., J. Urol. 163(2): 503-5120 (2000); Polascik et al., J. Urol. August; 162(2):293-306 (1999) and Fortier et al., J. Nat. Cancer Inst. 91(19): 1635-1640 (1999)). A variety of other diagnostic markers are also used in similar contexts including p53 and K-ras (see, e.g., Tulchinsky et al., Int J Mol Med 1999 Jul. 4(1):99-102 and Minimoto et al., Cancer Detect Prev 2000; 24(1):1-12). Therefore, this disclosure of 98P4B6 polynucleotides and polypeptides (as well as 98P4B6 polynucleotide probes and anti-98P4B6 antibodies used to identify the presence of these molecules) and their properties allows skilled artisans to utilize these molecules in methods that are analogous to those used, for example, in a variety of diagnostic assays directed to examining conditions associated with cancer.

Typical embodiments of diagnostic methods which utilize the 98P4B6 polynucleotides, polypeptides, reactive T cells and antibodies are analogous to those methods from well-established diagnostic assays, which employ, e.g., PSA polynucleotides, polypeptides, reactive T cells and antibodies. For example, just as PSA polynucleotides are used as probes (for example in Northern analysis, see, e.g., Sharief et al., Biochem. Mol. Biol. 1 mL 33(3):567-74 (1994)) and primers (for example in PCR analysis, see, e.g., Okegawa et al., J. Urol. 163(4): 1189-1190 (2000)) to observe the presence and/or the level of PSA mRNAs in methods of monitoring PSA overexpression or the metastasis of prostate cancers, the 98P4B6 polynucleotides described herein can be utilized in the same way to detect 98P4B6 overexpression or the metastasis of prostate and other cancers expressing this gene. Alternatively, just as PSA polypeptides are used to generate antibodies specific for PSA which can then be used to observe the presence and/or the level of PSA proteins in methods to monitor PSA protein overexpression (see, e.g., Stephan et al., Urology 55(4):560-3 (2000)) or the metastasis of prostate cells (see, e.g., Alanen et al., Pathol. Res. Pract. 192(3):233-7 (1996)), the 98P4B6 polypeptides described herein can be utilized to generate antibodies for use in detecting 98P4B6 overexoression or the metastasis of prostate cells and cells of other cancers expressing this gene.

Specifically, because metastases involves the movement of cancer cells from an organ of origin (such as the lung or prostate gland etc.) to a different area of the body (such as a lymph node), assays which examine a biological sample for the presence of cells expressing 98P4B6 polynucleotides and/or polypeptides can be used to provide evidence of metastasis. For example, when a biological sample from tissue that does not normally contain 98P4B6-expressing cells (lymph node) is found to contain 98P4B6-expressing cells such as the 98P4B6 expression seen in LAPC4 and LAPC9, xenografts isolated from lymph node and bone metastasis, respectively, this finding is indicative of metastasis.

Alternatively 98P4B6 polynucleotides and/or polypeptides can be used to provide evidence of cancer, for example, when cells in a biological sample that do not normally express 98P4B6 or express 98P4B6 at a different level are found to express 98P4B6 or have an increased expression of 98P4B6 (see, e.g., the 98P4B6 expression in the cancers listed in Table I and in patient samples etc. shown in the accompanying Figures). In such assays, artisans may further wish to generate supplementary evidence of metastasis by testing the biological sample for the presence of a second tissue restricted marker (in addition to 98P4B6) such as PSA, PSCA etc. (see, e.g., Alanen et al., Pathol. Res. Pract. 192(3): 233-237 (1996)).

Just as PSA polynucleotide fragments and polynucleotide variants are employed by skilled artisans for use in methods of monitoring PSA, 98P4B6 polynucleotide fragments and polynucleotide variants are used in an analogous manner. In particular, typical PSA polynucleotides used in methods of monitoring PSA are probes or primers which consist of fragments of the PSA cDNA sequence. Illustrating this, primers used to PCR amplify a PSA polynucleotide must include less than the whole PSA sequence to function in the polymerase chain reaction. In the context of such PCR reactions, skilled artisans generally create a variety of different polynucleotide fragments that can be used as primers in order to amplify different portions of a polynucleotide of interest or to optimize amplification reactions (see, e.g., Caetano-Anolles, G. Biotechniques 25(3): 472-476, 478-480 (1998); Robertson et al., Methods Mol. Biol. 98:121-154 (1998)). An additional illustration of the use of such fragments is provided in the Example entitled “Expression analysis of 98P4B6 in normal tissues, and patient specimens,” where a 98P4B6 polynucleotide fragment is used as a probe to show the expression of 98P4B6 RNAs in cancer cells. In addition, variant polynucleotide sequences are typically used as primers and probes for the corresponding mRNAs in PCR and Northern analyses (see, e.g., Sawai et al., Fetal Diagn. Ther. 1996 Nov.-Dec. 11(6):407-13 and Current Protocols In Molecular Biology, Volume 2, Unit 2, Frederick M. Ausubel et al. eds., 1995)). Polynucleotide fragments and variants are useful in this context where they are capable of binding to a target polynucleotide sequence (e.g., a 98P4B6 polynucleotide shown in FIG. 2 or variant thereof) under conditions of high stringency.

Furthermore, PSA polypeptides which contain an epitope that can be recognized by an antibody or T cell that specifically binds to that epitope are used in methods of monitoring PSA. 98P4B6 polypeptide fragments and polypeptide analogs or variants can also be used in an analogous manner. This practice of using polypeptide fragments or polypeptide variants to generate antibodies (such as ant-PSA antibodies or T cells) is typical in the art with a wide variety of systems such as fusion proteins being used by practitioners (see, e.g., Current Protocols In Molecular Biology, Volume 2, Unit 16, Frederick M. Ausubel et al. eds., 1995). In this context, each epitope(s) functions to provide the architecture with which an antibody or T cell is reactive. Typically, skilled artisans create a variety of different polypeptide fragments that can be used in order to generate immune responses specific for different portions of a polypeptide of interest (see, e.g., U.S. Pat. No. 5,840,501 and U.S. Pat. No. 5,939,533). For example it may be preferable to utilize a polypeptide comprising one of the 98P4B6 biological motifs discussed herein or a motif-bearing subsequence which is readily identified by one of skill in the art based on motifs available in the art. Polypeptide fragments, variants or analogs are typically useful in this context as long as they comprise an epitope capable of generating an antibody or T cell specific for a target polypeptide sequence (e.g. a 98P4B6 polypeptide shown in FIG. 3).

As shown herein, the 98P4B6 polynucleotides and polypeptides (as well as the 98P4B6 polynucleotide probes and anti-98P4B6 antibodies or T cells used to identify the presence of these molecules) exhibit specific properties that make them useful in diagnosing cancers such as those listed in Table I. Diagnostic assays that measure the presence of 98P4B6 gene products, in order to evaluate the presence or onset of a disease condition described herein, such as prostate cancer, are used to identify patients for preventive measures or further monitoring, as has been done so successfully with PSA. Moreover, these materials satisfy a need in the art for molecules having similar or complementary characteristics to PSA in situations where, for example, a definite diagnosis of metastasis of prostatic origin cannot be made on the basis of a test for PSA alone (see, e.g., Alanen et al., Pathol. Res. Pract. 192(3): 233-237 (1996)), and consequently, materials such as 98P4B6 polynucleotides and polypeptides (as well as the 98P4B6 polynucleotide probes and anti-98P4B6 antibodies used to identify the presence of these molecules) need to be employed to confirm a metastases of prostatic origin.

Finally, in addition to their use in diagnostic assays, the 98P4B6 polynucleotides disclosed herein have a number of other utilities such as their use in the identification of oncogenetic associated chromosomal abnormalities in the chromosomal region to which the 98P4B6 gene maps (see the Example entitled “Chromosomal Mapping of 98P4B6” below). Moreover, in addition to their use in diagnostic assays, the 98P4B6-related proteins and polynucleotides disclosed herein have other utilities such as their use in the forensic analysis of tissues of unknown origin (see, e.g., Takahama K Forensic Sci Int 1996 Jun. 28; 80(1-2): 63-9).

Additionally, 98P4B6-related proteins or polynucleotides of the invention can be used to treat a pathologic condition characterized by the over-expression of 98P4B6. For example, the amino acid or nucleic acid sequence of FIG. 2 or FIG. 3, or fragments of either, can be used to generate an immune response to a 98P4B6 antigen. Antibodies or other molecules that react with 98P4B6 can be used to modulate the function of this molecule, and thereby provide a therapeutic benefit.

XII.) INHIBITION OF 98P4B6 PROTEIN FUNCTION

The invention includes various methods and compositions for inhibiting the binding of 98P4B6 to its binding partner or its association with other protein(s) as well as methods for inhibiting 98P4B6 function.

XII.A.) Inhibition of 98P4B6 with Intracellular Antibodies

In one approach, a recombinant vector that encodes single chain antibodies that specifically bind to 98P4B6 are introduced into 98P4B6 expressing cells via gene transfer technologies. Accordingly, the encoded single chain anti-98P4B6 antibody is expressed intracellularly, binds to 98P4B6 protein, and thereby inhibits its function. Methods for engineering such intracellular single chain antibodies are well known. Such intracellular antibodies, also known as “intrabodies”, are specifically targeted to a particular compartment within the cell, providing control over where the inhibitory activity of the treatment is focused. This technology has been successfully applied in the art (for review, see Richardson and Marasco, 1995, TIBTECH vol. 13). Intrabodies have been shown to virtually eliminate the expression of otherwise abundant cell surface receptors (see, e.g., Richardson et al., 1995, Proc. Natl. Acad. Sci. USA 92: 3137-3141; Beerli et al., 1994, J. Biol. Chem. 289: 23931-23936; Deshane et al., 1994, Gene Ther. 1: 332-337).

Single chain antibodies comprise the variable domains of the heavy and light chain joined by a flexible linker polypeptide, and are expressed as a single polypeptide. Optionally, single chain antibodies are expressed as a single chain variable region fragment joined to the light chain constant region. Well-known intracellular trafficking signals are engineered into recombinant polynucleotide vectors encoding such single chain antibodies in order to target precisely the intrabody to the desired intracellular compartment. For example, intrabodies targeted to the endoplasmic reticulum (ER) are engineered to incorporate a leader peptide and, optionally, a C-terminal ER retention signal, such as the KDEL amino acid motif. Intrabodies intended to exert activity in the nucleus are engineered to include a nuclear localization signal. Lipid moieties are joined to intrabodies in order to tether the intrabody to the cytosolic side of the plasma membrane. Intrabodies can also be targeted to exert function in the cytosol. For example, cytosolic intrabodies are used to sequester factors within the cytosol, thereby preventing them from being transported to their natural cellular destination.

In one embodiment, intrabodies are used to capture 98P4B6 in the nucleus, thereby preventing its activity within the nucleus. Nuclear targeting signals are engineered into such 98P4B6 intrabodies in order to achieve the desired targeting. Such 98P4B6 intrabodies are designed to bind specifically to a particular 98P4B6 domain. In another embodiment, cytosolic intrabodies that specifically bind to a 98P4B6 protein are used to prevent 98P4B6 from gaining access to the nucleus, thereby preventing it from exerting any biological activity within the nucleus (e.g., preventing 98P4B6 from forming transcription complexes with other factors).

In order to specifically direct the expression of such intrabodies to particular cells, the transcription of the intrabody is placed under the regulatory control of an appropriate tumor-specific promoter and/or enhancer. In order to target intrabody expression specifically to prostate, for example, the PSA promoter and/or promoter/enhancer can be utilized (See, for example, U.S. Pat. No. 5,919,652 issued 6 Jul. 1999).

XII.B.) Inhibition of 98P4B6 with Recombinant Proteins

In another approach, recombinant molecules bind to 98P4B6 and thereby inhibit 98P4B6 function. For example, these recombinant molecules prevent or inhibit 98P4B6 from accessing/binding to its binding partner(s) or associating with other protein(s). Such recombinant molecules can, for example, contain the reactive part(s) of a 98P4B6 specific antibody molecule. In a particular embodiment, the 98P4B6 binding domain of a 98P4B6 binding partner is engineered into a dimeric fusion protein, whereby the fusion protein comprises two 98P4B6 ligand binding domains linked to the Fc portion of a human IgG, such as human IgG1. Such IgG portion can contain, for example, the C_(H)2 and C_(H)3 domains and the hinge region, but not the C_(H)1 domain. Such dimeric fusion proteins are administered in soluble form to patients suffering from a cancer associated with the expression of 98P4B6, whereby the dimeric fusion protein specifically binds to 98P4B6 and blocks 98P4B6 interaction with a binding partner. Such dimeric fusion proteins are further combined into multimeric proteins using known antibody linking technologies.

XII.C.) Inhibition of 98P4B6 Transcription or Translation

The present invention also comprises various methods and compositions for inhibiting the transcription of the 98P4B6 gene. Similarly, the invention also provides methods and compositions for inhibiting the translation of 98P4B6 mRNA into protein.

In one approach, a method of inhibiting the transcription of the 98P4B6 gene comprises contacting the 98P4B6 gene with a 98P4B6 antisense polynucleotide. In another approach, a method of inhibiting 98P4B6 mRNA translation comprises contacting a 98P4B6 mRNA with an antisense polynucleotide. In another approach, a 98P4B6 specific ribozyme is used to cleave a 98P4B6 message, thereby inhibiting translation. Such antisense and ribozyme based methods can also be directed to the regulatory regions of the 98P4B6 gene, such as 98P4B6 promoter and/or enhancer elements. Similarly, proteins capable of inhibiting a 98P4B6 gene transcription factor are used to inhibit 98P4B6 mRNA transcription. The various polynucleotides and compositions useful in the aforementioned methods have been described above. The use of antisense and ribozyme molecules to inhibit transcription and translation is well known in the art.

Other factors that inhibit the transcription of 98P4B6 by interfering with 98P4B6 transcriptional activation are also useful to treat cancers expressing 98P4B6. Similarly, factors that interfere with 98P4B6 processing are useful to treat cancers that express 98P4B6. Cancer treatment methods utilizing such factors are also within the scope of the invention.

XII.D.) General Considerations for Therapeutic Strategies

Gene transfer and gene therapy technologies can be used to deliver therapeutic polynucleotide molecules to tumor cells synthesizing 98P4B6 (i.e., antisense, ribozyme, polynucleotides encoding intrabodies and other 98P4B6 inhibitory molecules). A number of gene therapy approaches are known in the art Recombinant vectors encoding 98P4B6 antisense polynucleotides, ribozymes, factors capable of interfering with 98P4B6 transcription, and so forth, can be delivered to target tumor cells using such gene therapy approaches.

The above therapeutic approaches can be combined with any one of a wide variety of surgical, chemotherapy or radiation therapy regimens. The therapeutic approaches of the invention can enable the use of reduced dosages of chemotherapy (or other therapies) and/or less frequent administration, an advantage for all patients and particularly for those that do not tolerate the toxicity of the chemotherapeutic agent well.

The anti-tumor activity of a particular composition (e.g., antisense, ribozyme, intrabody), or a combination of such compositions, can be evaluated using various in vitro and in vivo assay systems. In vitro assays that evaluate therapeutic activity include cell growth assays, soft agar assays and other assays indicative of tumor promoting activity, binding assays capable of determining the extent to which a therapeutic composition will inhibit the binding of 98P4B6 to a binding partner, etc.

In vivo, the effect of a 98P4B6 therapeutic composition can be evaluated in a suitable animal model. For example, xenogeneic prostate cancer models can be used, wherein human prostate cancer explants or passaged xenograft tissues are introduced into immune compromised animals, such as nude or SCID mice (Klein et al., 1997, Nature Medicine 3: 402-408). For example, PCT Patent Application WO98/16628 and U.S. Pat. No. 6,107,540 describe various xenograft models of human prostate cancer capable of recapitulating the development of primary tumors, micrometastasis, and the formation of osteoblastic metastases characteristic of late stage disease. Efficacy can be predicted using assays that measure inhibition of tumor formation, tumor regression or metastasis, and the like.

In vivo assays that evaluate the promotion of apoptosis are useful in evaluating therapeutic compositions. In one embodiment, xenografts from tumor bearing mice treated with the therapeutic composition can be examined for the presence of apoptotic foci and compared to untreated control xenograft-bearing mice. The extent to which apoptotic foci are found in the tumors of the treated mice provides an indication of the therapeutic efficacy of the composition.

The therapeutic compositions used in the practice of the foregoing methods can be formulated into pharmaceutical compositions comprising a carrier suitable for the desired delivery method. Suitable carriers include any material that when combined with the therapeutic composition retains the anti-tumor function of the therapeutic composition and is generally non-reactive with the patient's immune system. Examples include, but are not limited to, any of a number of standard pharmaceutical carriers such as sterile phosphate buffered saline solutions, bacteriostatic water, and the like (see, generally, Remington's Pharmaceutical Sciences 16^(th) Edition, A. Osal., Ed., 1980).

Therapeutic formulations can be solubilized and administered via any route capable of delivering the therapeutic composition to the tumor site. Potentially effective routes of administration include, but are not limited to, intravenous, parenteral, intraperitoneal, intramuscular, intratumor, intradermal, intraorgan, orthotopic, and the like. A preferred formulation for intravenous injection comprises the therapeutic composition in a solution of preserved bacteriostatic water, sterile unpreserved water, and/or diluted in polyvinylchloride or polyethylene bags containing 0.9% sterile Sodium Chloride for Injection, USP. Therapeutic protein preparations can be lyophilized and stored as sterile powders, preferably under vacuum, and then reconstituted in bacteriostatic water (containing for example, benzyl alcohol preservative) or in sterile water prior to injection.

Dosages and administration protocols for the treatment of cancers using the foregoing methods will vary with the method and the target cancer, and will generally depend on a number of other factors appreciated in the art.

XIII.) IDENTIFICATION, CHARACTERIZATION AND USE OF MODULATORS OF 98P4B6

Methods to Identify and Use Modulators

In one embodiment, screening is performed to identify modulators that induce or suppress a particular expression profile, suppress or induce specific pathways, preferably generating the associated phenotype thereby. In another embodiment, having identified differentially expressed genes important in a particular state; screens are performed to identify modulators that alter expression of individual genes, either increase or decrease. In another embodiment, screening is performed to identify modulators that alter a biological function of the expression product of a differentially expressed gene. Again, having identified the importance of a gene in a particular state, screens are performed to identify agents that bind and/or modulate the biological activity of the gene product.

In addition, screens are done for genes that are induced in response to a candidate agent. After identifying a modulator (one that suppresses a cancer expression pattern leading to a normal expression pattern, or a modulator of a cancer gene that leads to expression of the gene as in normal tissue) a screen is performed to identify genes that are specifically modulated in response to the agent. Comparing expression profiles between normal tissue and agent-treated cancer tissue reveals genes that are not expressed in normal tissue or cancer tissue, but are expressed in agent treated tissue, and vice versa. These agent-specific sequences are identified and used by methods described herein for cancer genes or proteins. In particular these sequences and the proteins they encode are used in marking or identifying agent-treated cells. In addition, antibodies are raised against the agent-induced proteins and used to target novel therapeutics to the treated cancer tissue sample.

Modulator-Related Identification and Screening Assays:

Gene Expression-Related Assays

Proteins, nucleic acids, and antibodies of the invention are used in screening assays. The cancer-associated proteins, antibodies, nucleic adds, modified proteins and cells containing these sequences are used in screening assays, such as evaluating the effect of drug candidates on a “gene expression profile,” expression profile of polypeptides or alteration of biological function. In one embodiment, the expression profiles are used, preferably in conjunction with high throughput screening techniques to allow monitoring for expression profile genes after treatment with a candidate agent (e.g., Davis, G F, et al., J Biol Screen 7:69 (2002); Zlokarnik, et al., Science 279:84-8 (1998); Heid, Genome Res 6:986-94, 1996).

The cancer proteins, antibodies, nucleic acids, modified proteins and cells containing the native or modified cancer proteins or genes are used in screening assays. That is, the present invention comprises methods for screening for compositions which modulate the cancer phenotype or a physiological function of a cancer protein of the invention. This is done on a gene itself or by evaluating the effect of drug candidates on a “gene expression profile” or biological function. In one embodiment, expression profiles are used, preferably in conjunction with high throughput screening techniques to allow monitoring after treatment with a candidate agent, see Zlokarnik, supra.

A variety of assays are executed directed to the genes and proteins of the invention. Assays are run on an individual nucleic acid or protein level. That is, having identified a particular gene as up regulated in cancer, test compounds are screened for the ability to modulate gene expression or for binding to the cancer protein of the invention. “Modulation” in this context includes an increase or a decrease in gene expression. The preferred amount of modulation will depend on the original change of the gene expression in normal versus tissue undergoing cancer, with changes of at least 10%, preferably 50%, more preferably 100-300%, and in some embodiments 300-1000% or greater. Thus, if a gene exhibits a 4-fold increase in cancer tissue compared to normal tissue, a decrease of about four-fold is often desired; similarly, a 10-fold decrease in cancer tissue compared to normal tissue a target value of a 10-fold increase in expression by the test compound is often desired. Modulators that exacerbate the type of gene expression seen in cancer are also useful, e.g., as an upregulated target in further analyses.

The amount of gene expression is monitored using nucleic acid probes and the quantification of gene expression levels, or, alternatively; a gene product itself is monitored, e.g., through the use of antibodies to the cancer protein and standard immunoassays. Proteomics and separation techniques also allow for quantification of expression.

Expression Monitoring to Identify Compounds that Modify Gene Expression

In one embodiment, gene expression monitoring, i.e., an expression profile, is monitored simultaneously for a number of entities. Such profiles will typically involve one or more of the genes of FIG. 2. In this embodiment, e.g., cancer nucleic acid probes are attached to biochips to detect and quantify cancer sequences in a particular cell. Alternatively, PCR can be used. Thus, a series, e.g., wells of a microtiter plate, can be used with dispensed primers in desired wells. A PCR reaction can then be performed and analyzed for each well.

Expression monitoring is performed to identify compounds that modify the expression of one or more cancer-associated sequences, e.g., a polynucleotide sequence set out in FIG. 2. Generally, a test modulator is added to the cells prior to analysis. Moreover, screens are also provided to identify agents that modulate cancer, modulate cancer proteins of the invention, bind to a cancer protein of the invention, or interfere with the binding of a cancer protein of the invention and an antibody or other binding partner.

In one embodiment, high throughput screening methods involve providing a library containing a large number of potential therapeutic compounds (candidate compounds). Such “combinatorial chemical libraries” are then screened in one or more assays to identify those library members (particular chemical species or subclasses) that display a desired characteristic activity. The compounds thus identified can serve as conventional “lead compounds,” as compounds for screening, or as therapeutics.

In certain embodiments, combinatorial libraries of potential modulators are screened for an ability to bind to a cancer polypeptide or to modulate activity. Conventionally, new chemical entities with useful properties are generated by identifying a chemical compound (called a “lead compound”) with some desirable property or activity, e.g., inhibiting activity, creating variants of the lead compound, and evaluating the property and activity of those variant compounds. Often, high throughput screening (HTS) methods are employed for such an analysis.

As noted above, gene expression monitoring is conveniently used to test candidate modulators (e.g., protein, nucleic acid or small molecule). After the candidate agent has been added and the cells allowed to incubate for a period, the sample containing a target sequence to be analyzed is, e.g., added to a biochip.

If required, the target sequence is prepared using known techniques. For example, a sample is treated to lyse the cells, using known lysis buffers, electroporation, etc., with purification and/or amplification such as PCR performed as appropriate. For example, an in vitro transcription with labels covalently attached to the nucleotides is performed. Generally, the nucleic acids are labeled with biotin-FITC or PE, or with cy3 or cy5.

The target sequence can be labeled with, e.g., a fluorescent, a chemiluminescent, a chemical, or a radioactive signal, to provide a means of detecting the target sequence's specific binding to a probe. The label also can be an enzyme, such as alkaline phosphatase or horseradish peroxidase, which when provided with an appropriate substrate produces a product that is detected. Alternatively, the label is a labeled compound or small molecule, such as an enzyme inhibitor, that binds but is not catalyzed or altered by the enzyme. The label also can be a moiety or compound, such as, an epitope tag or biotin which specifically binds to streptavidin. For the example of biotin, the streptavidin is labeled as described above, thereby, providing a detectable signal for the bound target sequence. Unbound labeled streptavidin is typically removed prior to analysis.

As will be appreciated by those in the art, these assays can be direct hybridization assays or can comprise “sandwich assays”, which include the use of multiple probes, as is generally outlined in U.S. Pat. Nos. 5,681,702; 5,597,909; 5,545,730; 5,594,117; 5,591,584; 5,571,670; 5,580,731; 5,571,670; 5,591,584; 5,624,802; 5,635,352; 5,594,118; 5,359,100; 5,124,246; and 5,681,697. In this embodiment, in general, the target nucleic acid is prepared as outlined above, and then added to the biochip comprising a plurality of nucleic acid probes, under conditions that allow the formation of a hybridization complex.

A variety of hybridization conditions are used in the present invention, including high, moderate and low stringency conditions as outlined above. The assays are generally run under stringency conditions which allow formation of the label probe hybridization complex only in the presence of target. Stringency can be controlled by altering a step parameter that is a thermodynamic variable, including, but not limited to, temperature, formamide concentration, salt concentration, chaotropic salt concentration pH, organic solvent concentration, etc. These parameters may also be used to control non-specific binding, as is generally outlined in U.S. Pat. No. 5,681,697. Thus, it can be desirable to perform certain steps at higher stringency conditions to reduce non-specific binding.

The reactions outlined herein can be accomplished in a variety of ways. Components of the reaction can be added simultaneously, or sequentially, indifferent orders, with preferred embodiments outlined below. In addition, the reaction may include a variety of other reagents. These include salts, buffers, neutral proteins, e.g. albumin, detergents, etc. which can be used to facilitate optimal hybridization and detection, and/or reduce nonspecific or background interactions. Reagents that otherwise improve the efficiency of the assay, such as protease inhibitors, nuclease inhibitors, anti-microbial agents, etc., may also be used as appropriate, depending on the sample preparation methods and purity of the target. The assay data are analyzed to determine the expression levels of individual genes, and changes in expression levels as between states, forming a gene expression profile.

Biological Activity-Related Assays

The invention provides methods identify or screen for a compound that modulates the activity of a cancer-related gene or protein of the invention. The methods comprise adding a test compound, as defined above, to a cell comprising a cancer protein of the invention. The cells contain a recombinant nucleic acid that encodes a cancer protein of the invention.

In another embodiment, a library of candidate agents is tested on a plurality of cells.

In one aspect, the assays are evaluated in the presence or absence or previous or subsequent exposure of physiological signals, e.g. hormones, antibodies, peptides, antigens, cytokines, growth factors, action potentials, pharmacological agents including chemotherapeutics, radiation, carcinogenics, or other cells (i.e., cell-cell contacts). In another example, the determinations are made at different stages of the cell cycle process. In this way, compounds that modulate genes or proteins of the invention are identified. Compounds with pharmacological activity are able to enhance or interfere with the activity of the cancer protein of the invention. Once identified, similar structures are evaluated to identify critical structural features of the compound.

In one embodiment, a method of modulating (e.g., inhibiting) cancer cell division is provided; the method comprises administration of a cancer modulator. In another embodiment, a method of modulating (e.g., inhibiting) cancer is provided; the method comprises administration of a cancer modulator. In a further embodiment, methods of treating cells or individuals with cancer are provided; the method comprises administration of a cancer modulator.

In one embodiment, a method for modulating the status of a cell that expresses a gene of the invention is provided. As used herein status comprises such art-accepted parameters such as growth, proliferation, survival, function, apoptosis, senescence, location, enzymatic activity, signal transduction, etc. of a cell. In one embodiment, a cancer inhibitor is an antibody as discussed above. In another embodiment, the cancer inhibitor is an antisense molecule. A variety of cell growth, proliferation, and metastasis assays are known to those of skill in the art, as described herein.

High Throughout Screening to Identify Modulators

The assays to identify suitable modulators are amenable to high throughput screening. Preferred assays thus detect enhancement or inhibition of cancer gene transcription, inhibition or enhancement of polypeptide expression, and inhibition or enhancement of polypeptide activity.

In one embodiment, modulators evaluated in high throughput screening methods are proteins, often naturally occurring proteins or fragments of naturally occurring proteins. Thus, e.g., cellular extracts containing proteins, or random or directed digests of proteinaceous cellular extracts, are used. In this way, libraries of proteins are made for screening in the methods of the invention. Particularly preferred in this embodiment are libraries of bacterial, fungal, viral, and mammalian proteins, with the latter being preferred, and human proteins being especially preferred. Particularly useful test compound will be directed to the class of proteins to which the target belongs, e.g., substrates for enzymes, or ligands and receptors.

Use of Soft Agar Growth and Colony Formation to Identify and Characterize Modulators

Normal cells require a solid substrate to attach and grow. When cells are transformed, they lose this phenotype and grow detached from the substrate. For example, transformed cells can grow in stirred suspension culture or suspended in semi-solid media, such as semi-solid or soft agar. The transformed-cells, when transfected with tumor suppressor genes, can regenerate normal phenotype and once again require a solid substrate to attach to and grow. Soft agar growth or colony formation in assays are used to identify modulators of cancer sequences, which when expressed in host cells, inhibit abnormal cellular proliferation and transformation. A modulator reduces or eliminates the host cells' ability to grow suspended in solid or semisolid media, such as agar.

Techniques for soft agar growth or colony formation in suspension assays are described in Freshney, Culture of Animal Cells a Manual of Basic Technique (3rd ed., 1994). See also, the methods section of Garkavtsev et al. (1996), supra.

Evaluation of Contact Inhibition and Growth Density Limitation to Identify and Characterize Modulators

Normal cells typically grow in a flat and organized pattern in cell culture until they touch other cells. When the cells touch one another, they are contact inhibited and stop growing. Transformed cells, however, are not contact inhibited and continue to grow to high densities in disorganized foci. Thus, transformed cells grow to a higher saturation density than corresponding normal cells. This is detected morphologically by the formation of a disoriented monolayer of cells or cells in foci. Alternatively, labeling index with (3H)-thymidine at saturation density is used to measure density limitation of growth, similarly an MTT or Alamar blue assay will reveal proliferation capacity of cells and the ability of modulators to affect same. See Freshney (1994), supra. Transformed cells, when transfected with tumor suppressor genes, can regenerate a normal phenotype and become contact inhibited and would grow to a lower density.

In this assay, labeling index with ³H)-thymidine at saturation density is a preferred method of measuring density limitation of growth. Transformed host cells are transfected with a cancer-associated sequence and are grown for 24 hours at saturation density in non-limiting medium conditions. The percentage of cells labeling with (3H)-thymidine is determined by incorporated cpm.

Contact independent growth is used to identify modulators of cancer sequences, which had led to abnormal cellular proliferation and transformation. A modulator reduces or eliminates contact independent growth, and returns the cells to a normal phenotype.

Evaluation of Growth Factor or Serum Dependence to Identify and Characterize Modulators

Transformed cells have lower serum dependence than their normal counterparts (see, e.g., Temin, J. Natl. Cancer Inst. 37:167-175 (1966); Eagle et al., J. Exp. Med. 131:836-879 (1970)); Freshney, supra. This is in part due to release of various growth factors by the transformed cells. The degree of growth factor or serum dependence of transformed host cells can be compared with that of control. For example, growth factor or serum dependence of a cell is monitored in methods to identify and characterize compounds that modulate cancer-associated sequences of the invention.

Use of Tumor-specific Marker Levels to Identify and Characterize Modulators Tumor cells release an increased amount of certain factors (hereinafter “tumor specific markers”) than their normal counterparts. For example, plasminogen activator (PA) is released from human glioma at a higher level than from normal brain cells (see, e.g., Gullino, Angiogenesis, Tumor Vascularization, and Potential Interference with Tumor Growth, in Biological Responses in Cancer, pp. 178-184 (Mihich (ed.) 1985)). Similarly, Tumor Angiogenesis Factor (TAF) is released at a higher level in tumor cells than their normal counterparts. See, e.g., Folkman, Angiogenesis and Cancer, Sem Cancer Biol. (1992)), while bFGF is released from endothelial tumors (Ensoli, B et al).

Various techniques which measure the release of these factors are described in Freshney (1994), supra. Also, see, Unkless et al., J. Biol. Chem. 249:4295-4305 (1974); Strickland & Beers, J. Biol. Chem. 251:5694-5702 (1976); Whur et al., Br. J. Cancer 42:305 312 (1980); Gullino, Angiogenesis, Tumor Vascularization, and Potential Interference with Tumor Growth, in Biological Responses in Cancer, pp. 178-184 (Mihich (ed.) 1985); Freshney, Anticancer Res. 5:111-130 (1985). For example, tumor specific marker levels are monitored in methods to identify and characterize compounds that modulate cancer-associated sequences of the invention.

Invasiveness into Matrigel to Identify and Characterize Modulators

The degree of invasiveness into Matrigel or an extracellular matrix constituent can be used as an assay to identify and characterize compounds that modulate cancer associated sequences. Tumor cells exhibit a positive correlation between malignancy and invasiveness of cells into Matrigel or some other extracellular matrix constituent. In this assay, tumorigenic cells are typically used as host cells. Expression of a tumor suppressor gene in these host cells would decrease invasiveness of the host cells. Techniques described in Cancer Res. 1999; 59:6010; Freshney (1994), supra, can be used. Briefly, the level of invasion of host cells is measured by using filters coated with Matrigel or some other extracellular matrix constituent. Penetration into the gel, or through to the distal side of the filter, is rated as invasiveness, and rated histologically by number of cells and distance moved, or by prelabeling the cells with ¹²⁵I and counting the radioactivity on the distal side of the filter or bottom of the dish. See, e.g., Freshney (1984), supra.

Evaluation of Tumor Growth In Vivo to Identify and Characterize Modulators

Effects of cancer-associated sequences on cell growth are tested in transgenic or immune-suppressed organisms. Transgenic organisms are prepared in a variety of art-accepted ways. For example, knock-out transgenic organisms, e.g., mammals such as mice, are made, in which a cancer gene is disrupted or in which a cancer gene is inserted. Knock-out transgenic mice are made by insertion of a marker gene or other heterologous gene into the endogenous cancer gene site in the mouse genome via homologous recombination. Such mice can also be made by substituting the endogenous cancer gene with a mutated version of the cancer gene, or by mutating the endogenous cancer gene, e.g., by exposure to carcinogens.

To prepare transgenic chimeric animals, e.g., mice, a DNA construct is introduced into the nuclei of embryonic stem cells. Cells containing the newly engineered genetic lesion are injected into a host mouse embryo, which is re-implanted into a recipient female. Some of these embryos develop into chimeric mice that possess germ cells some of which are derived from the mutant cell line. Therefore, by breeding the chimeric mice it is possible to obtain a new line of mice containing the introduced genetic lesion (see, e.g., Capecchi et al., Science 244:1288 (1989)). Chimeric mice can be derived according to U.S. Pat. No. 6,365,797, issued 2 Apr. 2002; U.S. Pat. No. 6,107,540 issued 22 Aug. 2000; Hogan et al., Manipulating the Mouse Embryo: A laboratory Manual, Cold Spring Harbor Laboratory (1988) and Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, Robertson, ed., IRL Press, Washington, D.C., (1987).

Alternatively, various immune-suppressed or immune-deficient host animals can be used. For example, a genetically athymic “nude” mouse (see, e.g., Giovanella et al., J. Natl. Cancer Inst. 52:921 (1974)), a SCID mouse, a thymectornized mouse, or an irradiated mouse (see, e.g., Bradley et al., Br. J. Cancer 38:263 (1978); Selby et al., Br. J. Cancer 41:52 (1980)) can be used as a host. Transplantable tumor cells (typically about 10⁶ cells) injected into isogenic hosts produce invasive tumors in a high proportion of cases, while normal cells of similar origin will not. In hosts which developed invasive tumors, cells expressing cancer-associated sequences are injected subcutaneously or orthotopically. Mice are then separated into groups, including control groups and treated experimental groups) e.g. treated with a modulator). After a suitable length of time, preferably 4-8 weeks, tumor growth is measured (e.g., by volume or by its two largest dimensions, or weight) and compared to the control. Tumors that have statistically significant reduction (using, e.g., Student's T test) are said to have inhibited growth.

In Vitro Assays to Identify and Characterize Modulators

Assays to identify compounds with modulating activity can be performed in vitro. For example, a cancer polypeptide is first contacted with a potential modulator and incubated for a suitable amount of time, e.g., from 0.5 to 48 hours. In one embodiment, the cancer polypeptide levels are determined in vitro by measuring the level of protein or mRNA. The level of protein is measured using immunoassays such as Western blotting, ELISA and the like with an antibody that selectively binds to the cancer polypeptide or a fragment thereof. For measurement of mRNA, amplification, e.g., using PCR, LCR, or hybridization assays, e.g., Northern hybridization, RNAse protection, dot blotting, are preferred. The level of protein or mRNA is detected using directly or indirectly labeled detection agents, e.g., fluorescently or radioactively labeled nucleic acids, radioactively or enzymatically labeled antibodies, and the like, as described herein.

Alternatively, a reporter gene system can be devised using a cancer protein promoter operably linked to a reporter gene such as luciferase, green fluorescent protein, CAT, or P-gal. The reporter construct is typically transfected into a cell. After treatment with a potential modulator, the amount of reporter gene transcription, translation, or activity is measured according to standard techniques known to those of skill in the art (Davis G F, supra; Gonzalez, J. & Negulescu, P. Curr. Opin. Biotechnol. 1998: 9:624).

As outlined above, in vitro screens are done on individual genes and gene products. That is, having identified a particular differentially expressed gene as important in a particular state, screening of modulators of the expression of the gene or the gene product itself is performed.

In one embodiment, screening for modulators of expression of specific gene(s) is performed. Typically, the expression of only one or a few genes is evaluated. In another embodiment, screens are designed to first find compounds that bind to differentially expressed proteins. These compounds are then evaluated for the ability to modulate differentially expressed activity. Moreover, once initial candidate compounds are identified, variants can be further screened to better evaluate structure activity relationships.

Binding Assays to Identify and Characterize Modulators

In binding assays in accordance with the invention, a purified or isolated gene product of the invention is generally used. For example, antibodies are generated to a protein of the invention, and immunoassays are run to determine the amount and/or location of protein. Alternatively, cells comprising the cancer proteins are used in the assays.

Thus, the methods comprise combining a cancer protein of the invention and a candidate compound such as a ligand, and determining the binding of the compound to the cancer protein of the invention. Preferred embodiments utilize the human cancer protein; animal models of human disease of can also be developed and used. Also, other analogous mammalian proteins also can be used as appreciated by those of skill in the art. Moreover, in some embodiments variant or derivative cancer proteins are used.

Generally, the cancer protein of the invention, or the ligand, is non-diffusibly bound to an insoluble support. The support can, e.g., be one having isolated sample receiving areas (a microtiter plate, an array, etc.). The insoluble supports can be made of any composition to which the compositions can be bound, is readily separated from soluble material, and is otherwise compatible with the overall method of screening. The surface of such supports can be solid or porous and of any convenient shape.

Examples of suitable insoluble supports include microtiter plates, arrays, membranes and beads. These are typically made of glass, plastic (e.g., polystyrene), polysaccharide, nylon, nitrocellulose, or Teflon™, etc. Microtiter plates and arrays are especially convenient because a large number of assays can be carried out simultaneously, using small amounts of reagents and samples. The particular manner of binding of the composition to the support is not crucial so long as it is compatible with the reagents and overall methods of the invention, maintains the activity of the composition and is nondiffusable. Preferred methods of binding include the use of antibodies which do not sterically block either the ligand binding site or activation sequence when attaching the protein to the support, direct binding to “sticky” or ionic supports, chemical crosslinking, the synthesis of the protein or agent on the surface, etc. Following binding of the protein or ligand/binding agent to the support, excess unbound material is removed by washing. The sample receiving areas may then be blocked through-incubation with bovine serum albumin (BSA), casein or other innocuous protein or other moiety.

Once a cancer protein of the invention is bound to the support, and a test compound is added to the assay. Alternatively, the candidate binding agent is bound to the support and the cancer protein of the invention is then added. Binding agents include specific antibodies, non-natural binding agents identified in screens of chemical libraries, peptide analogs, etc.

Of particular interest are assays to identify agents that have a low toxicity for human cells. A wide variety of assays can be used for this purpose, including proliferation assays, CAMP assays, labeled in vitro protein-protein binding assays, electrophoretic mobility shift assays, immunoassays for protein binding, functional assays (phosphorylation assays, etc.) and the like.

A determination of binding of the test compound (ligand, binding agent, modulator, etc.) to a cancer protein of the invention can be done in a number of ways. The test compound can be labeled, and binding determined directly, e.g., by attaching all or a portion of the cancer protein of the invention to a solid support, adding a labeled candidate compound (e.g., a fluorescent label), washing off excess reagent, and determining whether the label is present on the solid support. Various blocking and washing steps can be utilized as appropriate.

In certain embodiments, only one of the components is labeled, e.g., a protein of the invention or ligands labeled. Alternatively, more than one component is labeled with different labels, e.g., I¹²⁵, for the proteins and a fluorophor for the compound. Proximity reagents, e.g., quenching or energy transfer reagents are also useful.

Competitive Binding to Identify and Characterize Modulators

In one embodiment, the binding of the “test compound” is determined by competitive binding assay with a “competitor.” The competitor is a binding moiety that binds to the target molecule (e.g., a cancer protein of the invention). Competitors include compounds such as antibodies, peptides, binding partners, ligands, etc. Under certain circumstances, the competitive binding between the test compound and the competitor displaces the test compound. In one embodiment, the test compound is labeled. Either the test compound, the competitor, or both, is added to the protein for a time sufficient to allow binding. Incubations are performed at a temperature that facilitates optimal activity, typically between four and 40° C. Incubation periods are typically optimized, e.g., to facilitate rapid high throughput screening; typically between zero and one hour will be sufficient. Excess reagent is generally removed or washed away. The second component is then added, and the presence or absence of the labeled component is followed, to indicate binding.

In one embodiment, the competitor is added first, followed by the test compound. Displacement of the competitor is an indication that the test compound is binding to the cancer protein and thus is capable of binding to, and potentially modulating, the activity of the cancer protein. In this embodiment, either component can be labeled. Thus, e.g., if the competitor is labeled, the presence of label in the post-test compound wash solution indicates displacement by the test compound. Alternatively, if the test compound is labeled, the presence of the label on the support indicates displacement.

In an alternative embodiment, the test compound is added first, with incubation and washing, followed by the competitor. The absence of binding by the competitor indicates that the test compound binds to the cancer protein with higher affinity than the competitor. Thus, if the test compound is labeled, the presence of the label on the support, coupled with a lack of competitor binding, indicates that the test compound binds to and thus potentially modulates the cancer protein of the invention.

Accordingly, the competitive binding methods comprise differential screening to identity agents that are capable of modulating the activity of the cancer proteins of the invention. In this embodiment, the methods comprise combining a cancer protein and a competitor in a first sample. A second sample comprises a test compound, the cancer protein, and a competitor. The binding of the competitor is determined for both samples, and a change, or difference in binding between the two samples indicates the presence of an agent capable of binding to the cancer protein and potentially modulating its activity. That is, if the binding of the competitor is different in the second sample relative to the first sample, the agent is capable of binding to the cancer protein.

Alternatively, differential screening is used to identify drug candidates that bind to the native cancer protein, but cannot bind to modified cancer proteins. For example the structure of the cancer protein is modeled and used in rational drug design to synthesize agents that interact with that site, agents which generally do not bind to site-modified proteins. Moreover, such drug candidates that affect the activity of a native cancer protein are also identified by screening drugs for the ability to either enhance or reduce the activity of such proteins.

Positive controls and negative controls can be used in the assays. Preferably control and test samples are performed in at least triplicate to obtain statistically significant results. Incubation of all samples occurs for a time sufficient to allow for the binding of the agent to the protein. Following incubation, samples are washed free of non-specifically bound material and the amount of bound, generally labeled agent determined. For example, where a radiolabel is employed, the samples can be counted in a scintillation counter to determine the amount of bound compound.

A variety of other reagents can be included in the screening assays. These include reagents like salts, neutral proteins, e.g. albumin, detergents, etc. which are used to facilitate optimal protein-protein binding and/or reduce non-specific or background interactions. Also reagents that otherwise improve the efficiency of the assay, such as protease inhibitors, nuclease inhibitors, anti-microbial agents, etc., can be used. The mixture of components is added in an order that provides for the requisite binding.

Use of Polynucleotides to Down-regulate or Inhibit a Protein of the Invention.

Polynucleotide modulators of cancer can be introduced into a cell containing the target nucleotide sequence by formation of a conjugate with a ligand-binding molecule, as described in WO 91/04753. Suitable ligand-binding molecules include, but are not limited to, cell surface receptors, growth factors, other cytokines, or other ligands that bind to cell surface receptors. Preferably, conjugation of the ligand binding molecule does not substantially interfere with the ability of the ligand binding molecule to bind to its corresponding molecule or receptor, or block entry of the sense or antisense oligonucleotide or its conjugated version into the cell. Alternatively, a polynucleotide modulator of cancer can be introduced into a cell containing the target nucleic acid sequence, e.g., by formation of a polynucleotide-lipid complex, as described in WO 90/10448. It is understood that the use of antisense molecules or knock out and knock in models may also be used in screening assays as discussed above, in addition to methods of treatment.

Inhibitory and Antisense Nucleotides

In certain embodiments, the activity of a cancer-associated protein is down-regulated, or entirely inhibited, by the use of antisense polynucleotide or inhibitory small nuclear RNA (snRNA), i.e., a nucleic acid complementary to, and which can preferably hybridize specifically to, a coding mRNA nucleic acid sequence, e.g., a cancer protein of the invention, mRNA, or a subsequence thereof. Binding of the antisense polynucleotide to the mRNA reduces the translation and/or stability of the mRNA.

In the context of this invention, antisense polynucleotides can comprise naturally occurring nucleotides, or synthetic species formed from naturally occurring subunits or their close homologs. Antisense polynucleotides may also have altered sugar moieties or inter-sugar linkages. Exemplary among these are the phosphorothioate and other sulfur containing species which are known for use in the art. Analogs are comprised by this invention so long as they function effectively to hybridize with nucleotides of the invention. See, e.g., Isis Pharmaceuticals, Carlsbad, Calif.; Sequitor, Inc., Natick, Mass.

Such antisense polynucleotides can readily be synthesized using recombinant means, or can be synthesized in vitro. Equipment for such synthesis is sold by several vendors, including Applied Biosystems. The preparation of other oligonucleotides such as phosphorothioates and alkylated derivatives is also well known to those of skill in the art;

Antisense molecules as used herein include antisense or sense oligonucleotides. Sense oligonucleotides can, e.g., be employed to block transcription by binding to the anti-sense strand. The antisense and sense oligonucleotide comprise a single stranded nucleic acid sequence (either RNA or DNA) capable of binding to target mRNA (sense) or DNA (antisense) sequences for cancer molecules. Antisense or sense oligonucleotides, according to the present invention, comprise a fragment generally at least about 12 nucleotides, preferably from about 12 to 30 nucleotides. The ability to derive an antisense or a sense oligonucleotide, based upon a cDNA sequence encoding a given protein is described in, e.g., Stein &Cohen (Cancer Res. 48:2659 (1988 and van der Krol et al. (BioTechniques 6:958 (1988)).

Ribozymes

In addition to antisense polynucleotides, ribozymes can be used to target and inhibit transcription of cancer-associated nucleotide sequences. A ribozyme is an RNA molecule that catalytically cleaves other RNA molecules. Different kinds of ribozymes have been described, including group I ribozymes, hammerhead ribozymes, hairpin ribozymes, RNase P, and axhead ribozymes (see, e.g., Castanolto et al., Adv. in Pharmacology 25: 289-317 (1994) for a general review of the properties of different ribozymes).

The general features of hairpin ribozymes are described, e.g., in Hampel et al., Nucl. Acids Res. 18:299-304 (1990); European Patent Publication No. 0360257; U.S. Pat. No. 5,254,678. Methods of preparing are well known to those of skill in the art (see, e.g., WO 94/26877; Ojwang et al., Proc. Natl. Acad. Sci. USA 90:6340-6344 (1993); Yamada et al., Human Gene Therapy 1:3945 (1994); Leavitt et al., Proc. Natl. Acad. Sci. USA 92:699-703 (1995); Leavitt et al., Human Gene Therapy 5: 1151-120 (1994); and Yamada et al., Virology 205: 121-126 (1994)).

Use of Modulators in Phenotypic Screening

In one embodiment, a test compound is administered to a population of cancer cells, which have an associated cancer expression profile. By “administration” or “contacting” herein is meant that the modulator is added to the cells in such a manner as to allow the modulator to act upon the cell, whether by uptake and intracellular action, or by action at the cell surface. In some embodiments, a nucleic acid encoding a proteinaceous agent (i.e., a peptide) is put into a viral construct such as an adenoviral or retroviral construct, and added to the cell, such that expression of the peptide agent is accomplished, e.g., PCT US97/01019. Regulatable gene therapy systems can also be used. Once the modulator has been administered to the cells, the cells are washed if desired and are allowed to incubate under preferably physiological conditions for some period. The cells are then harvested and a new gene expression profile is generated. Thus, e.g., cancer tissue is screened for agents that modulate, e.g., induce or suppress, the cancer phenotype. A change in at least one gene, preferably many, of the expression profile indicates that the agent has an effect on cancer activity. Similarly, altering a biological function or a signaling pathway is indicative of modulator activity. By defining such a signature for the cancer phenotype, screens for new drugs that alter the phenotype are devised. With this approach, the drug target need not be known and need not be represented in the original gene/protein expression screening platform, nor does the level of transcript for the target protein need to change. The modulator inhibiting function will serve as a surrogate marker

As outlined above, screens are done to assess genes or gene products. That is, having identified a particular differentially expressed gene as important in a particular state, screening of modulators of either the expression of the gene or the gene product itself is performed.

Use of Modulators to Affect Peptides of the Invention

Measurements of cancer polypeptide activity, or of the cancer phenotype are performed using a variety of assays. For example, the effects of modulators upon the function of a cancer polypeptide(s) are measured by examining parameters described above. A physiological change that affects activity is used to assess the influence of a test compound on the polypeptides of this invention. When the functional outcomes are determined using intact cells or animals, a variety of effects can be assesses such as, in the case of a cancer associated with solid tumors, tumor growth, tumor metastasis, neovascularization, hormone release, transcriptional changes to both known and uncharacterized genetic markers (e.g., by Northern blots), changes in cell metabolism such as cell growth or pH changes, and changes in intracellular second messengers such as cGNIP.

Methods of Identifying Characterizing Cancer-associated Sequences

Expression of various gene sequences is correlated with cancer. Accordingly, disorders based on mutant or variant cancer genes are determined. In one embodiment, the invention provides methods for identifying cells containing variant cancer genes, e.g., determining the presence of, all or part, the sequence of at least one endogenous cancer gene in a cell. This is accomplished using any number of sequencing techniques. The invention comprises methods of identifying the cancer genotype of an individual, e.g., determining all or part of the sequence of at least one gene of the invention in the individual. This is generally done in at least one tissue of the individual, e.g., a tissue set forth in Table I, and may include the evaluation of a number of tissues or different samples of the same tissue. The method may include comparing the sequence of the sequenced gene to a known cancer gene, i.e., a wild-type gene to determine the presence of family members, homologies, mutations or variants. The sequence of all or part of the gene can then be compared to the sequence of a known cancer gene to determine if any differences exist. This is done using any number of known homology programs, such as BLAST, Bestfit, etc. The presence of a difference in the sequence between the cancer gene of the patient and the known cancer gene correlates with a disease state or a propensity for a disease state, as outlined herein.

In a preferred embodiment, the cancer genes are used as probes to determine the number of copies of the cancer gene in the genome. The cancer genes are used as probes to determine the chromosomal localization of the cancer genes. Information such as chromosomal localization finds use in providing a diagnosis or prognosis in particular when chromosomal abnormalities such as translocations, and the like are identified in the cancer gene locus.

XIV.) KITS/ARTICLES OF MANUFACTURE

For use in the diagnostic and therapeutic applications described herein, kits are also within the scope of the invention. Such kits can comprise a carrier, package or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in the method. For example, the container(s) can comprise a probe that is or can be detectably labeled. Such probe can be an antibody or polynucleotide specific for a FIG. 2-related protein or a FIG. 2 gene or message, respectively. Where the method utilizes nucleic acid hybridization to detect the target nucleic acid, the kit can also have containers containing nucleotide(s) for amplification of the target nucleic acid sequence and/or a container comprising a reporter-means, such as a biotin-binding protein, such as avidin or streptavidin, bound to a reporter molecule, such as an enzymatic, florescent, or radioisotope label. The kit can include all or part of the amino acid sequences in FIG. 2 or FIG. 3 or analogs thereof, or a nucleic acid molecules that encodes such amino acid sequences.

The kit of the invention will typically comprise the container described above and one or more other containers comprising materials desirable from a commercial and user standpoint, including buffers, diluents, filters, needles, syringes; carrier, package, container, vial and/or tube labels listing contents and/or instructions for use, and package inserts with instructions for use.

A label can be present on the container to indicate that the composition is used for a specific therapy or non-therapeutic application, such as a diagnostic or laboratory application, and can also indicate directions for either in vivo or in vitro use, such as those described herein. Directions and or other information can also be included on an insert(s) or label(s) which is included with or on the kit.

The terms “kit” and “article of manufacture” can be used as synonyms.

In another embodiment of the invention, an article(s) of manufacture containing compositions, such as amino acid sequence(s), small molecule(s), nucleic acid sequence(s), and/or antibody(s), e.g., materials useful for the diagnosis, prognosis, prophylaxis and/or treatment of neoplasias of tissues such as those set forth in Table I is provided. The article of manufacture typically comprises at least one container and at least one label. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers can be formed from a variety of materials such as glass or plastic. The container can hold amino acid sequence(s), small molecule(s), nucleic acid sequence(s), and/or antibody(s), in one embodiment the container holds a polynucleotide for use in examining the mRNA expression profile of a cell, together with reagents used for this purpose.

The container can alternatively hold a composition which is effective for treating, diagnosis, prognosing or prophylaxing a condition and can have a sterile access port (for example the container can be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). The active agents in the composition can be an antibody capable of specifically binding 98P4B6 and modulating the function of 98P486.

The label can be on or associated with the container. A label a can be on a container when letters, numbers or other characters forming the label are molded or etched into the container itself; a label can be associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert. The label can indicate that the composition is used for diagnosing, treating, prophylaxing or prognosing a condition, such as a neoplasia of a tissue set forth in Table I. The article of manufacture can further comprise a second container comprising a pharmaceutically-acceptable buffer, such as phosphate-buffered saline, Ringer's solution and/or dextrose solution. It can further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, stirrers, needles, syringes, and/or package inserts with indications and/or instructions for use.

EXAMPLES

Various aspects of the invention are further described and illustrated by way of the several examples that follow, none of which are intended to limit the scope of the invention.

Example 1 SSH-Generated Isolation of cDNA Fragment of the 98P4B6 Gene

To isolate genes that are over-expressed in prostate cancer we used the Suppression Subtractive Hybridization (SSH) procedure using cDNA derived from prostate tissues. The 98P4B6 SSH cDNA sequence was derived from normal prostate minus LAPC-4AD prostate xenograft cDNAs. The 98P4B6 cDNA was identified as highly expressed in prostate cancer.

Materials and Methods

Human Tissues:

The patient cancer and normal tissues were purchased from different sources such as the NDRI (Philadelphia, Pa.). mRNA for some normal tissues were purchased from Clontech, Palo Alto, Calif.

RNA Isolation:

Tissues were homogenized in Trizol reagent (Life Technologies, Gibco BRL) using 10 ml/g tissue isolate total RNA. Poly A RNA was purified from total RNA using Qiagen's Oligotex mRNA Mini and Midi kits. Total and mRNA were quantified by spectrophotometric analysis (O.D. 260/280 nm) and analyzed by gel electrophoresis.

Oligonucleotides:

The following HPLC purified oligonucleotides were used.

DPNCDN (cDNA synthesis primer): (SEQ ID NO: 101) 5′TTTTGATCAAGCTT303′ Adaptor 1: (SEQ ID NO: 102) 5′CTAATACGACTCACTATAGGGCTCGAGCGGCCGCCCGGGCAG3′ (SEQ ID NO: 103) 3′GGCCCGTCCTAG5′ Adaptor 2: (SEQ ID NO: 104) 5′GTAATACGACTCACTATAGGGCAGCGTGGTCGCGGCCGAG3′ (SEQ ID NO: 105) 3′CGGCTCCTAG5′ PCR primer 1: (SEQ ID NO: 106) 5′CTAATACGACTCACTATAGGGC3′ Nested primer (NP)1: (SEQ ID NO: 107) 5′TCGAGCGGCCGCCCGGGCAGGA3′ Nested primer (NP)2: (SEQ ID NO: 108) 5′AGCGTGGTCGCGGCCGAGGA3′

Suppression Subtractive Hybridization:

Suppression Subtractive Hybridization (SSH) was used to identify cDNAs corresponding to genes that may be differentially expressed in prostate cancer. The SSH reaction utilized cDNA from prostate cancer xenograft and normal tissues.

The gene 98P4B6 sequence was derived from normal prostate tissue minus prostate cancer xenograft LAPC-4AD cDNA subtraction. The SSH DNA sequence (FIG. 1) was identified.

The cDNA derived from LAPC-4AD was used as the source of the “driver” cDNA, while the cDNA from normal prostate was used as the source of the “tester” cDNA. Double stranded cDNAs corresponding to tester and driver cDNAs were synthesized from 2 μg of poly(A)+ RNA isolated from the relevant tissue, as described above, using CLONTECH's PCR-Select cDNA Subtraction Kit and 1 ng of oligonucleotide DPNCDN as primer. First- and second-strand synthesis were carried out as described in the Kit's user manual protocol (CLONTECH Protocol No. PT1117-1, Catalog No. K1804-1). The resulting cDNA was digested with Dpn II for 3 hrs at 37° C. Digested cDNA was extracted with phenol/chloroform (1:1) and ethanol precipitated.

Driver cDNA was generated by combining in a 1:1 ratio Dpn II digested cDNA from the relevant tissue source (see above) with digested cDNAs derived from normal tissue.

Tester cDNA was generated by diluting 1 μl of Dpn II digested cDNA from the relevant tissue source (see above) (400 ng) in 5 μl of water. The diluted cDNA (2 μl, 160 ng) was then ligated to 2 μl of Adaptor 1 and Adaptor 2 (10 μM), in separate ligation reactions, in a total volume of 10 μl at 16° C. overnight, using 400 u of T4 DNA ligase (CLONTECH). Ligation was terminated with 1 μl of 0.2 M EDTA and heating at 72° C. for 5 min.

The first hybridization was performed by adding 1.5 μl (600 ng) of driver cDNA to each of two tubes containing 1.5 μl (20 ng) Adaptor 1- and Adaptor 2-ligated tester cDNA. In a final volume of 4 μl, the samples were overlaid with mineral oil, denatured in an MJ Research thermal cycler at 98° C. for 1.5 minutes, and then were allowed to hybridize for 8 hrs at 68° C. The two hybridizations were then mixed together with an additional 1 μl of fresh denatured driver cDNA and were allowed to hybridize overnight at 68° C. The second hybridization was then diluted in 200 μl of 20 mM Hepes, pH 8.3, 50 mM NaCl, 0.2 mM EDTA, heated at 70° C. for 7 min. and stored at −20° C.

PCR Amplification, Cloning and Sequencing of Gene Fragments Generated from SSH:

To amplify gene fragments resulting from SSH reactions, two PCR amplifications were performed. In the primary PCR reaction 1 μl of the diluted final hybridization mix was added to 1 μl of PCR primer 1 (10 μM), 0.5 μl dNTP mix (10 μM), 2.5 μl 10× reaction buffer (CLONTECH) and 0.5 μl 50× Advantage cDNA polymerase Mix (CLONTECH) in a final volume of 25 μl. PCR 1 was conducted using the following conditions: 75° C. for 5 min., 94° C. for 25 sec., then 27 cycles of 94° C. for 10 sec, 66° C. for 30 sec, 72° C. for 1.5 min. Five separate primary PCR reactions were performed for each experiment. The products were pooled and diluted 1:10 with water. For the secondary PCR reaction, 1 μl from the pooled and diluted primary PCR reaction was added to the same reaction mix as used for PCR 1, except that primers NP1 and NP2 (10 μM) were used instead of PCR primer 1. PCR 2 was performed using 10-12 cycles of 94° C. for 10 sec, 68° C. for 30 sec, and 72° C. for 1.5 minutes. The PCR products were analyzed using 2% agarose gel electrophoresis.

The PCR products were inserted into pCR2.1 using the T/A vector cloning kit (Invitrogen). Transformed E. coli were subjected to blue/white and ampicillin selection. White colonies were picked and arrayed into 96 well plates and were grown in liquid culture overnight. To identify inserts, PCR amplification was performed on 1 ul of bacterial culture using the conditions of PCR1 and NP1 and NP2 as primers. PCR products were analyzed using 2% agarose gel electrophoresis.

Bacterial clones were stored in 20% glycerol in a 96 well format. Plasmid DNA was prepared, sequenced, and subjected to nucleic acid homology searches of the GenBank, dBest, and NCI-CGAP databases.

RT-PCR Expression Analysis:

First strand cDNAs can be generated from 1 μg of mRNA with oligo (dT)₁₋₂-18 priming using the Gibco-BRL Superscript Preamplification system. The manufacturer's protocol was used which included an incubation for 50 min at 42° C. with reverse transcriptase followed by RNAse H treatment at 37° C. for 20 min. After completing the reaction, the volume can be increased to 200 μl with water prior to normalization. First strand cDNAs from 16 different normal human tissues can be obtained from Clontech.

Normalization of the first strand cDNAs from multiple tissues was performed by using the primers 5′atatcgccgcgctcgtcgtcgacaa3′ (SEQ-ID NO: 109) and 5′gccacacgcagctcaftgtagaagg 3′ (SEQ ID NO: 110) to amplify β-actin. First strand cDNA (5 μl) were amplified in a total volume of 50 μl containing 0.4 μl primers, 0.2 μM each dNTPs, 1×PCR buffer (Clontech, 10 mM Tris-HCL, 1.5 mM MgCl₂, 50 mM KCl, pH8.3) and 1× Klentaq DNA polymerase (Clontech). Five μl of the PCR reaction can be removed at 18, 20, and 22 cycles and used for agarose gel electrophoresis. PCR was performed using an MJ Research thermal cycler under the following conditions: Initial denaturation can be at 94° C. for 15 sec, followed by a 18, 20, and 22 cycles of 94° C. for 15, 65° C. for 2 min, 72° C. for 5 sec. A final extension at 72° C. was carried out for 2 min. After agarose gel electrophoresis, the band intensifies of the 283 bp β-actin bands from multiple tissues were compared by visual inspection. Dilution factors for the first strand cDNAs were calculated to result in equal β-acfin band intensities in all tissues after 22 cycles of PCR. Three rounds of normalization can be required to achieve equal band intensifies in all tissues after 22 cycles of PCR.

To determine expression levels of the 98P4B6 gene, 5 μl of normalized first strand cDNA were analyzed by PCR using 26, and 30 cycles of amplification. Semi-quantitative expression analysis can be achieved by comparing the PCR products at cycle numbers that give light band intensities. The primers used for RT-PCR were designed using the 98P4B6 SSH sequence and are listed below:

98P4B6.1 5′- GACTGAGCTGGAACTGGAATTTGT- 3′ (SEQ ID NO: 111) 98P4B6.2 5′- TTTGAGGAGACTTCATCTCACTGG -3′ (SEQ ID NO: 112)

Example 2 Isolation of Full Length 98P4B6 Encoding cDNA

The 98P4B6 SSH cDNA sequence was derived from a substraction consisting of normal prostate minus prostate cancer xenograft. The SSH cDNA sequence (FIG. 1) was designated 98P4B6.

The 98P4B6 SSH DNA sequence of 183 bp is shown in FIG. 1. Full-length 98P4B6 v.1 (done GTD3) of 2453 bp was cloned from prostate cDNA library, revealing an ORF of 454 amino acids (FIG. 2 and FIG. 3). 98P4B6 v.6 was also cloned from normal prostate library. Other variants of 98P4B6 were also identified and these are listed in FIGS. 2 and 3.

98P4B6 v.2, v.3, v.4, v.5, v.6, v.7 and v.8 are splice variants of 98P4B6 v.1. 98P4B6 v.9 through v.19 are SNP variants and differ from v.1 by one amino acid. 98P4B6 v.20 through v.24 are SNP variants of v.7. 98P4B6 v.25 through v.38 are SNP variants of v.8. Though these SNP variants were shown separately, they could also occur in any combinations and in any transcript variants.

Example 3 Chromosomal Mapping of 98P4B6

Chromosomal localization can implicate genes in disease pathogenesis. Several chromosome mapping approaches are available including fluorescent in situ hybridization (FISH), human/hamster radiation hybrid (R H) panels (Walter et al., 1994; Nature Genetics 7:22; Research Genetics, Huntsville Ala.), human-rodent somatic cell hybrid panels such as is available from the Cornell Institute (Camden, N.J.), and genomic viewers utilizing BLAST homologies to sequenced and mapped genomic clones (NCBI, Bethesda, Md.).

98P4B6 maps to chromosome 7q21 using 98P4B6 sequence and the NCBI BLAST tool.

Example 4 Expression Analysis of 98P4B6

Expression analysis by RT-PCR demonstrated that 98P4B6 is strongly expressed in prostate cancer patient specimens (FIG. 14). First strand cDNA was generated from normal stomach, normal brain, normal heart, normal liver, normal skeletal muscle, normal testis, normal prostate, normal bladder, normal kidney, normal colon, normal lung, normal pancreas, and a pool of cancer specimens from prostate cancer patients, bladder cancer patients, kidney cancer patients, colon cancer patients, lung cancer patients, pancreas cancer patients, and a pool of 2 patient prostate metastasis to lymph node. Normalization was performed by PCR using primers to actin. Semi-quantitative PCR, using primers directed to 98P4B6 v.1, v.13, or/and v.14 (A), or directed specifically to the splice variants 98P4B6 v.6 and v.8 (B), was performed at 26 and 30 cycles of amplification. Samples were run on an agarose gel, and PCR products were quantitated using the AlphaImager software. Results show strong expression of 98P4B6 and its splice variants v.6 and v.8 in normal prostate and in prostate cancer. Expression was also detected in bladder cancer, kidney cancer, colon cancer, lung cancer, pancreas cancer, breast cancer, cancer metastasis as well as in the prostate cancer metastasis to lymph node specimens, compared to all normal tissues tested. As noted below, e.g., in Example 6, as 98P4B6 v.1 is in expressed in cancer tissues such as those listed in Table 1, the other protein-encoding 98P4B6 variants are expressed in these tissues as well; this principle is corroborated by data in (FIG. 14) for the proteins herein designated 98P4B6 v.6 or v.8 is found, e.g., in prostate, lung, ovary, bladder, breast, colon, kidney and pancreas, cancers, as well as in the literature (Porkka et al., Lab Invest, 2002 and Korkmaz et. al., JBC, 2002) where the protein 98P4B6 v.8 is identified in normal prostate and prostate cancer.

When the genomic region to which a gene maps is modulated in a particular cancer, the alternative transcripts or splice variants of the gene are modulated as well. Disclosed herein is that 98P4B6 has a particular expression profile related to cancer. Alternative transcripts and splice variants of 98P4B6 are also involved in cancers in the same or additional tissues, thus serving as tumor-associated markers/antigens.

Expression of 98P4B6 v.1, v.13, and/or v.14 was detected in prostate, lung, ovary, bladder, cervix, uterus and pancreas cancer patient specimens (FIG. 15). First strand cDNA was prepared from a panel of patient cancer specimens. Normalization was performed by PCR using primers to actin. Semi-quantitative PCR, using primers to 98P4B6, was performed at 26 and 30 cycles of amplification. Samples were run on an agarose gel, and PCR products were quantitated using the AlphaImager software. Expression was recorded as absent, low, medium or strong. Results show expression of 98P4B6 in the majority of all patent cancer specimens tested.

FIG. 16 shows that 98P4B6 is expressed in stomach cancer patient specimens. (A) RNA was extracted from normal stomach (N) and from 10 different stomach cancer patient specimens (T). Northern blot with 10 μg of total RNA/lane was probed with 98P4B6 sequence. Results show strong expression of 98P4B6 in the stomach tumor tissues and lower expression in normal stomach. The lower panel represents ethidium bromide staining of the blot showing quality of the RNA samples. (B) Expression of 98P4B6 was assayed in a panel of human stomach cancers (T) and their respective matched normal tissues (N) on RNA dot blots. 98P4B6 was detected in 7 out of 8 stomach tumors but not in the matched normal tissues.

Example 5 Transcript Variants of 98P4B6

Transcript variants are variants of mature mRNA from the same gene which arise by alternative transcription or alternative splicing. Alternative transcripts are transcripts from the same gene but start transcription at different points. Splice variants are mRNA variants spliced differently from the same transcript. In eukaryotes, when a multi-exon gene is transcribed from genomic DNA, the initial RNA is spliced to produce functional mRNA, which has only exons and is used for translation into an amino acid sequence. Accordingly, a given gene can have zero to many alternative transcripts and each transcript can have zero to many splice variants. Each transcript variant has a unique exon makeup, and can have different coding and/or non-coding (5′ or 3′ end) portions, from the original transcript. Transcript variants can code for similar or different proteins with the same or a similar function or can encode proteins with different functions, and can be expressed in the same tissue at the same time or in different tissues at the same time or in the same issue at different times or in different tissues at different times. Proteins encoded by transcript variants can have similar or different cellular or extracellular localizations, e.g., secreted versus intracellular.

Transcript variants are identified by a variety of art-accepted methods. For example, alternative transcripts and splice variants are identified by full-length cloning experiment, or by use of full-length transcript and EST sequences. First, all human ESTs were grouped into clusters which show direct or indirect identity with each other. Second, ESTs in the same cluster were further grouped into sub-clusters and assembled into a consensus sequence. The original gene sequence is compared to the consensus sequence(s) or other full-length sequences. Each consensus sequence is a potential splice variant for that gene. Even when a variant is identified that is not a full-length clone, that portion of the variant is very useful for antigen generation and for further cloning of the full-length splice variant, using techniques known in the art.

Moreover, computer programs are available in the art that identify transcript variants based on genomic sequences. Genomic-based transcript variant identification programs include FgenesH (A. Salamov and V. Solovyev, “Ab initoo gene finding in Drosophila genomic DNA,” Genome Research. 2000 April; 10(4):516-22); Grail and GenScan. For a general discussion of splice variant identification protocols see., e.g., Southan, C., A genomic perspective on human proteases, FEBS Lett. 2001 Jun. 8; 498(2-3):214-8; de Souza, S. J., et al., Identification of human chromosome 22 transcribed sequences with ORF expressed sequence tags, Proc. Natl. Acad Sci USA. 2000 Nov. 7; 97(23):12690-3.

To further confirm the parameters of a transcript variant, a variety of techniques are available in the art, such as full-length cloning, proteomic validation, PCR-based validation, and 5′ RACE validation, etc. (see e.g., Proteomic Validation: Brennan, S. O., et al., Albumin banks peninsula: a new termination variant characterized by electrospray mass spectrometry, Biochem Biophys Acta. 1999 Aug. 17; 1433(1-2):321-6; Ferranti P, et al., Differential splicing of pre-messenger RNA produces multiple forms of mature caprine alpha(s1)-casein, Eur J Biochem. 1997 Oct. 1; 249(1):1-7. For PCR-based Validation: Wellmann S, et al., Specific reverse transcription-PCR quantification of vascular endothelial growth factor (VEGF) splice variants by LightCycler technology, Clin Chem. 2001 April; 47(4):654-60; Jia, H. P., et al., Discovery of new human beta-defensins using a genomics-based approach, Gene. 2001 Jan. 24; 263(1-2):211-8. For PCR-based and 5′ RACE Validation: Brigle, K. E., et al., Organization of the murine reduced folate carrier gene and identification of variant splice forms, Biochem Biophys Acta. 1997 Aug. 7; 1353(2):191-8).

It is known in the art that genomic regions are modulated in cancers. Recently, Porkka et al. (2002) reported that transcript variants of STEAP2 were expressed and were found in both normal and malignant prostate tissue (Porkka, K. P., et al. Cloning and characterization of a novel six-transmembrane protein STEAP2, expressed in normal and malignant prostate. Laboratory Investigation 2002 November; 82(11):1573-1582). Another group of scientists also reported that transcript variants of STEAP2 (98P4B6 v.6 herein) also were expressed significantly higher in prostate cancer than normal prostate (Korkmaz, K. S., et al. Molecular cloning and characterization of STAMP1, a highly prostate-specific six transmembrane protein that is overexpressed in prostate cancer. The Journal of Biological Chemistry. 2002 September 277(39):36689-36696.). When the genomic region to which a gene maps is modulated in a particular cancer, the alternative transcripts or splice variants of the gene are modulated as well. Disclosed herein is that 98P4B6 has a particular expression profile related to cancer. Alternative transcripts and splice variants of 98P4B6 are also involved in cancers in the same or additional tissues, thus serving as tumor-associated markers/antigens.

Using the full-length gene and EST sequences, seven transcript variants were identified, designated as 98P4B6 v.2, v.3, v.4, v.5, v.6, v.7 and v.8, as shown in FIG. 12. The boundaries of exons in the original transcript, 98P4B6 v.1 were shown in Table LI. The first 22 bases of v.1 were not in the nearby 5′ region of v.1 on the current assembly of the human genome. Compared with 98P4B6 v.1, variant v.2 was a single exon transcript whose 3′ portion was the same as the last exon of v.1. The first two exons of v.3 were in intron 1 of v. 1. Variants v.4, v.5, and v.6 spliced out 224-334 in the first exon of v.1. In addition, v.5 spliced out exon 5 while v.6 spliced out exon 6 but extended exon 5 of v.1. Variant v.7 used alternative transcription start and different 3′ exons. Variant v.8 extended 5′ end and kept the whole intron 5 of v.1. Theoretically, each different combination of exons in spatial order, e.g. exons 2 and 3, is a potential splice variant.

Tables LII through LV are set forth on a variant-by-variant basis. Tables LII(a)-(g) show the nucleotide sequence of the transcript variant. Tables LII(a)-(g) show the alignment of the transcript variant with the nucleic acid sequence of 98P4B6 v.1. Tables LIV(a)-(g) lay out the amino acid translation of the transcript variant for the identified reading frame orientation. Tables LV(a)-(g) display alignments of the amino acid sequence encoded by the splice variant with that of 98P4B6 v.1. Additionally, single nucleotide polymorphisms (SNP) are noted in the alignment.

Example 6 Single Nucleotide Polymorphisms of 98P4B6

A Single Nucleotide Polymorphism (SNP) is a single base pair variation in a nucleotide sequence at a specific location. At any given point of the genome, there are four possible nucleotide base pairs: A/T, C/G, G/C and T/A. Genotype refers to the specific base pair sequence of one or more locations in the genome of an individual. Haplotype refers to the base pair sequence of more than one location on the same DNA molecule (or the same chromosome in higher organisms), often in the context of one gene or in the context of several tightly linked genes. SNP that occurs on a cDNA is called cSNP. This cSNP may change amino acids of the protein encoded by the gene and thus change the functions of the protein. Some SNP cause inherited diseases; others contribute to quantitative variations in phenotype and reactions to environmental factors including diet and drugs among individuals. Therefore, SNP and/or combinations of alleles (called haplotypes) have many applications, including diagnosis of inherited diseases, determination of drug reactions and dosage, identification of genes responsible for diseases, and analysis of the genetic relationship between individuals (P. Nowotny, J. M. Kwon and A. M. Goate, “SNP analysis to dissect human traits,” Curr. Opin. Neurobiol. 2001 October; 11(5):637-641; M. Pirmohamed and B. K. Park, “Genetic susceptibility to adverse drug reactions,” Trends Pharmacol. Sci. 2001 June; 22(6):298-305; J. H. Riley, C. J. Allan, E. Lai and A. Roses, “The use of single nucleotide polymorphisms in the isolation of common disease genes,” Pharmacogenomics. 2000 February; 1(1):39-47; R. Judson, J. C. Stephens and A. Windemuth, “The predictive power of haplotypes in clinical response,” Pharmacogenomics. 2000 feb; 1(1):15-26).

SNP are identified by a variety of art-accepted methods (P. Bean, “The promising voyage of SNP target discovery,” Am. Clin. Lab. 2001 October-November; 20(9):18-20; K. M. Weiss, “In search of human variation,” Genome Res. 1998 July; 8(7):691-697; M. M. She, “Enabling large-scale pharmacogenetic studies by high-throughput mutation detection and genotyping technologies,” Clin. Chem. 2001 February; 47(2):164-172). For example, SNP can be identified by sequencing DNA fragments that show polymorphism by gel-based methods such as restriction fragment length polymorphism (RFLP) and denaturing gradient gel electrophoresis (DGGE). They can also be discovered by direct sequencing of DNA samples pooled from different individuals or by comparing sequences from different DNA samples. With the rapid accumulation of sequence data in public and private databases, one can discover SNP by comparing sequences using computer programs (Z. Gu, L. Hillier and P. Y. Kwok, “Single nucleotide polymorphism hunting in cyberspace,” Hum. Mutat. 1998; 12(4):221-225). SNP can be verified and genotype or haplotype of an individual can be determined by a variety of methods including direct sequencing and high throughput microarrays (P. Y. Kwok, “Methods for genotyping single nucleotide polymorphisms,” Annu. Rev. Genomics Hum. Genet. 2001; 2:235-258; M. Kokoris, K. Dix, K. Moynihan, J. Mathis, B. Erwin, P. Grass, B. Hines and A. Duesterhoeft, “High-throughput SNP genotyping with the Masscode system,” Mol. Diagn. 2000 December; 5(4):329-340).

Using the methods described above, eleven SNP were identified in the original transcript, 98P4B6 v.1, at positions 46 (AIG), 179 (CfT), 180 (A/G), 269 (A/G), 404 (G/T), 985 (CfT), 1170 (T/C), 1497 (ANG), 1746 (T/G), 2046 (T/G) and 2103 (T/C). The transcripts or proteins with alternative allele were designated as variant 98P4B6 v.9 through v.19, as shown in FIG. 10 a. FIG. 11 shows the schematic alignment of protein variants, corresponding to nucleotide variants. Nucleotide variants that code for the same amino acid sequence as v.1 are not shown in FIG. 11. These alleles of the SNP, though shown separately here, can occur in different combinations (haplotypes) and in any one of the transcript variants (such as 98P4B6 v.5) that contains the site of the SNP. In addition, there were SNP in other transcript variants in regions not shared with v.1. For example, there were fourteen SNP in the fifth intron of v.1, which was part of transcript variants v.2, v.6 and v.8. These SNP are shown in FIG. 10 c and listed as following (numbers relative v.8): 1760 (G/A), 1818 (G/T), 1870 (C/T), 2612 (T/C), 2926 (T/A), 4241 (T/A), 4337 (AIG), 4338 (A/C), 4501 (A/G), 4506 (C/T), 5434 (C/A), 5434 (C/G), 5434 (C/T) and 5589 (C/A). FIG. 10 b shows the SNP in the unique regions of transcript variant v.7: 1956 (A/C), 1987 (T/A), 2010 (G/C), 2010 (G/T) and 2059 (G/A) (numbers correspond to nucleotide sequence of v.7).

Example 7 Production of Recombinant 98P4B6 in Prokaryotic Systems

To express recombinant 98P4B6 and 98P4B6 variants in prokaryotic cells, the full or partial length 98P4B6 and 98P4B6 variant cDNA sequences are cloned into any one of a variety of expression vectors known in the art. One or more of the following regions of 98P4B6 variants are expressed: the full length sequence presented in FIGS. 2 and 3, or any 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more contiguous amino acids from 98P4B6, variants, or analogs thereof.

A. In Vitro Transcription and Translation Constructs:

pCRII: To generate 98P4B6 sense and anti-sense RNA probes for RNA in situ investigations, pCRII constructs (Invitrogen, Carlsbad Calif.) are generated encoding either all or fragments of the 98P4B6 cDNA. The pCRII vector has Sp6 and T7 promoters flanking the insert to drive the transcription of 98P4B6 RNA for use as probes in RNA in situ hybridization experiments. These probes are used to analyze the cell and tissue expression of 98P4B6 at the RNA level. Transcribed 98P4B6 RNA representing the cDNA amino acid coding region of the 98P4B6 gene is used in in vitro translation systems such as the TnT™ Coupled Reticulolysate System (Promega, Corp., Madison, Wis.) to synthesize 98P4B6 protein.

B. Bacterial Constructs:

pGEX Constructs: To generate recombinant 98P4B6 proteins in bacteria that are fused to the Glutathione S-transferase (GST) protein, all or parts of the 98P4B6 cDNA protein coding sequence are cloned into the pGEX family of GST-fusion vectors (Amersham Pharmacia Biotech, Piscataway, N.J.). These constructs allow controlled expression of recombinant 98P4B6 protein sequences with GST fused at the amino-terminus and a six histidine epitope (6× His) at the carboxyl-terminus. The GST and 6× His tags permit purification of the recombinant fusion protein from induced bacteria with the appropriate affinity matrix and allow recognition of the fusion protein with anti-GST and anti-His antibodies. The 6× His tag is generated by adding 6 histidine codons to the cloning primer at the 3′ end, e.g., of the open reading frame (ORF). A proteolytic cleavage site, such as the PreScission™ recognition site in pGEX-6P-1, may be employed such that it permits cleavage of the GST tag from 98P4B6-related protein. The ampicillin resistance gene and pBR322 origin permits selection and maintenance of the pGEX plasmids in E. coli. A glutathione-5-transferase (GST) fusion protein encompassing amino acids 2-204 of the STEAP-2.protein sequence was generated in the pGEX vector. The recombinant GST-STEAP-2 fusion protein was purified from induced bacteria by glutathione-sepaharose affinity chromatography and used as immunogen for generation of a polyclonal antibody.

pMAL Constructs: To generate, in bacteria, recombinant 98P4B6 proteins that are fused to maltose-binding protein (MBP), all or parts of the 98P4B6 cDNA protein coding sequence are fused to the MBP gene by cloning into the pMAL-c2X and pMAL-p2X vectors (New England Biolabs, Beverly, Mass.). These constructs allow controlled expression of recombinant 98P4B6 protein sequences with MBP fused at the amino-terminus and a 6× His epitope tag at the carboxyl-terminus. The MBP and 6× His tags permit purification of the recombinant protein from induced bacteria with the appropriate affinity matrix and allow recognition of the fusion protein with anti-MBP and anti-His antibodies. The 6× His epitope tag is generated by adding 6 histidine codons to the 3′ cloning primer. A Factor Xa recognition site permits cleavage of the pMAL tag from 98P4B6. The pMAL-c2X and pMAL-p2X vectors are optimized to express the recombinant protein in the cytoplasm or periplasm respectively. Periplasm expression enhances folding of proteins with disulfide bonds.

pET Constructs: To express 98P4B6 in bacterial cells, all or parts of the 98P4B6 cDNA protein coding sequence are cloned into the pET family of vectors (Novagen, Madison, Wis.). These vectors allow tightly controlled expression of recombinant 98P4B6 protein in bacteria with and without fusion to proteins that enhance solubility, such as NusA and thioredoxin (Trx), and epitope tags, such as 6× His and S-Tag™ that aid purification and detection of the recombinant protein. For example, constructs are made utilizing pET NusA fusion system 43.1 such that regions of the 98P4B6 protein are expressed as amino-terminal fusions to NusA.

C. Yeast Constructs:

pESC Constructs: To express 98P4B6 in the yeast species Saccharomyces cerevisiae for generation of recombinant protein and functional studies, all or parts of the 98P4B6 cDNA protein coding sequence are cloned into the pESC family of vectors each of which contain 1 of 4 selectable markers, HIS3, TRP1, LEU2, and URA3 (Stratagene, La Jolla, Calif.). These vectors allow controlled expression from the same plasmid of up to 2 different genes or cloned sequences containing either Flag™ or Myc epitope tags in the same yeast cell. This system is useful to confirm protein-protein interactions of 98P4B6. In addition, expression in yeast yields similar post-translational modifications, such as glycosylations and phosphorylations, that are found when expressed in eukaryotic cells.

pESP Constructs: To express 98P4B6 in the yeast species Saccharomyces pombe, all or parts of the 98P4B6 cDNA protein coding sequence are cloned into the pESP family of vectors. These vectors allow controlled high level of expression of a 98P4B6 protein sequence that is fused at either the amino terminus or at the carboxyl terminus to GST which aids purification of the recombinant protein. A Flag™ epitope tag allows detection of the recombinant protein with anti-Flag™ antibody.

Example 8 Production of Recombinant 98P4B6 in Higher Eukaryotic Systems

A. Mammalian Constructs:

To express recombinant 98P4B6 in eukaryotic cells, the full or partial length 98P4B6 cDNA sequences can be cloned into any one of a variety of expression vectors known in the art. One or more of the following regions of 98P4B6 are expressed in these constructs, amino adds 1 to 255, or any 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more contiguous amino adds from 98P4B6 v.1 through v.11; amino adds 1 to 1266, or any 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more contiguous amino adds from 98P4B6 v.12 and v.13, variants, or analogs thereof.

The constructs can be transfected into any one of a wide variety of mammalian cells such as 293T cells. Transfected 293T cell lysates can be probed with the anti-98P4B6 polyclonal serum, described herein.

pcDNA4/HisMax Constructs: To express 98P4B6 in mammalian cells, a 98P4B6 ORF, or portions thereof, of 98P4B6 are cloned into pcDNA4/HisMax Version A (Invitrogen, Carlsbad, Calif.). Protein expression is driven from the cytomegalovirus (CMV) promoter and the SP16 translational enhancer. The recombinant protein has Xpress™ and six histidine (6× His) epitopes fused to the amino-terminus. The pcDNA4 MHisMax vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Zeocin resistance gene allows for selection of mammalian cells expressing the protein and the ampicillin resistance gene and ColE1 origin permits selection and maintenance of the plasmid in E. coli.

pcDNA3.1/MycHis Constructs: To express 98P4B6 in mammalian cells, a 98P4B6 ORF, or portions thereof, of 98P4B6 with a consensus Kozak translation initiation site was cloned into pcDNA3.1/MycHis Version A (Invitrogen, Carlsbad, Calif.). Protein expression is driven from the cytomegalovirus (CMV) promoter. The recombinant proteins have the myc epitope and 6× His epitope fused to the carboxyl-terminus. The pcDNA3.1/MycHis vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability, along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Neomycin resistance gene can be used, as it allows for selection of mammalian cells expressing the protein and the ampicillin resistance gene and ColE1 origin permits selection and maintenance of the plasmid in E. coli. pcDNA3.1/GFP Construct: To express 98P4B6 in mammalian cells and to allow detection of the recombinant proteins using fluorescence, the 98P4B6 ORF sequence was codon optimized according to Mirzabekov et al. (1999), and was cloned into pcDNA3.1/GFP vector to generate 98P4B6.GFP.pcDNA3.1 construct. Protein expression was driven from the cytomegalovirus (CMV) promoter. The recombinant protein had the Green Fluorescent Protein (GFP) fused to the carboxyl-terminus facilitating non-invasive, in vivo detection and cell biology studies. The pcDNA3.1/GFP vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Neomycin resistance gene allows for selection of mammalian cells that express the protein, and the ampicillin resistance gene and ColE1 origin permits selection and maintenance of the plasmid in E. coli.

Transfection of 98P4B6.GFP.pcDNA3.1 into 293T cells was performed as shown in FIGS. 17 and 18. Results show strong expression of the fusion protein by western blot analysis (FIG. 17), flow cytometry (FIG. 18A) and fluorescent microscopy (FIG. 18B).

Additional constructs with an amino-terminal GFP fusion are made in pcDNA3.1/NT-GFP-TOPO spanning the entire length of a 98P4B6 protein.

PAPtag: A 98P4B6 ORF, or portions thereof, is cloned into pAPtag-5 (GenHunter Corp. Nashville, Tenn.). This construct generates an alkaline phosphatase fusion at the carboxyl-terminus of a 98P4B6 protein while fusing the IgGκ signal sequence to the amino-terminus. Constructs are also generated in which alkaline phosphatase with an amino-terminal IgGκ signal sequence is fused to the amino-terminus of a 98P4B6 protein. The resulting recombinant 98P4B6 proteins are optimized for secretion into the media of transfected mammalian cells and can be used to identify proteins such as ligands or receptors that interact with 98P4B6 proteins. Protein expression is driven from the CMV promoter and the recombinant proteins also contain myc and 6× His epitopes fused at the carboxyl-terminus that facilitates detection and purification. The Zeocin resistance gene present in the vector allows for selection of mammalian cells expressing the recombinant protein and the ampicillin resistance gene permits selection of the plasmid in E. coli.

pTag5: A 98P4B6 ORF, or portions thereof, is cloned into pTag-5. This vector is similar to pAPtag but without the alkaline phosphatase fusion. This construct generates 98P4B6 protein with an amino-terminal IgGκ signal sequence and myc and 6× His epitope tags at the carboxyl-terminus that facilitate detection and affinity purification. The resulting recombinant 98P4B6 protein is optimized for secretion into the media of transfected mammalian cells, and is used as immunogen or ligand to identify proteins such as ligands or receptors that interact with the 98P4B6 proteins. Protein expression is driven from the CMV promoter. The Zeocin resistance gene present in the vector allows for selection of mammalian cells expressing the protein, and the ampicillin resistance gene permits selection of the plasmid in E. coli.

PsecFc: A 98P4B6 ORF, or portions thereof, is also cloned into psecFc. The psecFc vector was assembled by cloning the human immunoglobulin G1 (IgG) Fc (hinge, CH2, CH3 regions) into pSecTag2 (Invitrogen, California). This construct generates an IgG1 Fc fusion at the carboxyl-terminus of the 98P4B6 proteins, while fusing the IgGκ signal sequence to N-terminus. 98P4B6 fusions utilizing the murine IgG1 Fc region are also used. The resulting recombinant 98P4B6 proteins are optimized for secretion into the media of transfected mammalian cells, and can be used as immunogens or to identify proteins such as ligands or receptors that interact with 98P4B6 protein. Protein expression is driven from the CMV promoter. The hygromycin resistance gene present in the vector allows for selection of mammalian cells that express the recombinant protein, and the ampicillin resistance gene permits selection of the plasmid in E. coli.

pSRα Constructs: To generate mammalian cell lines that express 98P4B6 constitutively, 98P4B6 ORF, or portions thereof, of 98P4B6 were cloned into pSkα constructs. Amphotropic and ecotropic retroviruses were generated by transfecfion of pSRα constructs into the 293T-10A1 packaging line or co-transfection of pSRα and a helper plasmid (containing deleted packaging sequences) into the 293 cells, respectively. The retrovirus is used to infect a variety of mammalian cell lines, resulting in the integration of the cloned gene, 98P4B6, into the host cell-lines. Protein expression is driven from a long terminal repeat (LTR). The Neomycin resistance gene present in the vector allows for selection of mammalian cells that express the protein, and the ampicillin resistance gene and ColE1 origin permit selection and maintenance of the plasmid in E. coli. The retroviral vectors can thereafter be used for infection and generation of various cell lines using, for example, PC3, NIH 3T3, TsuPr1, 293 or rat-1 cells.

Additional pSRα constructs are made that fuse an epitope tag such as the FLAG™ tag to the carboxyl-terminus of 98P4B6 sequences to allow detection using anti-Flag antibodies. For example, the FLAG™ sequence 5′ gat tac aag gat gac gac gat aag 3′ (SEQ ID NO: 113) is added to cloning primer at the 3′ end of the ORF. Additional pSRα constructs are made to produce both amino-terminal and carboxyl-terminal GFP and myc/6× His fusion proteins of the full-length 98P4B6 proteins.

Additional Viral Vectors: Additional constructs are made for viral-mediated delivery and expression of 98P4B6. High virus filter leading to high level expression of 98P4B6 is achieved in viral delivery systems such as adenoviral vectors and herpes amplicon vectors. A 98P4B6 coding sequences or fragments thereof are amplified by PCR and subcloned into the AdEasy shuttle vector (Stratagene). Recombination and virus packaging are performed according to the manufacturer's instructions to generate adenoviral vectors. Alternatively, 98P4B6 coding sequences or fragments thereof are cloned into the HSV-1 vector (Imgenex) to generate herpes viral vectors. The viral vectors are thereafter used for infection of various cell lines such as PC3, NIH 3T3, 293 or rat-1 cells.

Regulated Expression Systems: To control expression of 98P4B6 in mammalian cells, coding sequences of 98P4B6, or portions thereof, are cloned into regulated mammalian expression systems such as the T-Rex System (Invitrogen), the GeneSwitch System (Invitrogen) and the tightly-regulated Ecdysone System (Sratagene). These systems allow the study of the temporal and concentration dependent effects of recombinant 98P4B6. These vectors are thereafter used to control expression of 98P4B6 in various cell lines such as PC3, NIH 3T3, 293 or rat-1 cells.

B. Baculovirus Expression Systems

To generate recombinant 98P4B6 proteins in a baculovirus expression system, 98P4B6 ORF, or portions thereof, are cloned into the baculovirus transfer vector pBlueBac 4.5 (Invitrogen), which provides a His-tag at the N-terminus. Specifically, pBlueBac-98P4B6 is co-transfected with helper plasmid pBac-N-Blue (Invitrogen) into SF9 (Spodoptera frugiperda) insect cells to generate recombinant baculovirus (see Invitrogen instruction manual for details). Baculovirus is then collected from cell supernatant and purified by plaque assay.

Recombinant 98P4B6 protein is then generated by infection of HighFive insect cells (Invitrogen) with purified baculovirus. Recombinant 98P4B6 protein can be detected using anti-98P4B6 or anti-His-tag antibody. 98P4B6 protein can be purified and used in various cell-based assays or as immunogen to generate polyclonal and monoclonal antibodies specific for 98P4B6.

Example 9 Antigenicity Profiles and Secondary Structure

FIG. 5(A-E), FIG. 6(A-E), FIG. 7(A-E), FIG. 8(A-E), and FIG. 9(A-E) depict graphically five amino acid profiles of 98P4B6 variants 1, 2, 5-7, each assessment available by accessing the ProtScale website located on the ExPasy molecular biology server.

These profiles: FIG. 5, Hydrophilicity, (Hopp T. P., Woods K. R., 1981. Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828); FIG. 6, Hydropathicity, (Kyte J., Doolittle R. F., 1982. J. Mol. Biol. 157:105-132); FIG. 7, Percentage Accessible Residues (Janin J., 1979 Nature 277:491-492); FIG. 8, Average Flexibility, (Bhaskaran R., and Ponnuswamy P. K., 1988. Int. J. Pept. Protein Res. 32:242-255); FIG. 9, Beta-turn (Deleage, G., Roux B. 1987 Protein Engineering 1:289-294); and optionally others available in the art, such as on the ProtScale website, were used to identify antigenic regions of each of the 98P4B6 variant proteins. Each of the above amino acid profiles of 98P4B6 variants were generated using the following ProtScale parameters for analysis: 1) A window size of 9; 2) 100% weight of the window edges compared to the window center; and, 3) amino acid profile values normalized to lie between 0 and 1.

Hydrophilicity (FIG. 5), Hydropathicity (FIG. 6) and Percentage Accessible Residues (FIG. 7) profiles were used to determine stretches of hydrophilic amino acids (i.e., values greater than 0.5 on the Hydrophilicity and Percentage Accessible Residues profile, and values less than 0.5 on the Hydropathicity profile). Such regions are likely to be exposed to the aqueous environment, be present on the surface of the protein, and thus available for immune recognition, such as by antibodies.

Average Flexibility (FIG. 8) and Beta-turn (FIG. 9) profiles determine stretches of amino acids (i.e., values greater than 0.5 on the Beta-turn profile and the Average Flexibility profile) that are not constrained in secondary structures such as beta sheets and alpha helices. Such regions are also more likely to be exposed on the protein and thus accessible to immune recognition, such as by antibodies.

Antigenic sequences of the 98P4B6 variant proteins indicated, e.g., by the profiles set forth in FIG. 5(A-E), FIG. 6(A-E), FIG. 7(A-E), FIG. 8(A-E), and/or FIG. 9(A-E) are used to prepare immunogens, either peptides or nucleic acids that encode them, to generate therapeutic and diagnostic anti-98P4B6 antibodies. The immunogen can be any 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50 or more than 50 contiguous amino acids, or the corresponding nucleic adds that encode them, from the 98P4B6 protein variants 1, 2, 5-7 listed in FIGS. 2 and 3. In particular, peptide immunogens of the invention can comprise, a peptide region of at least 5 amino adds of FIGS. 2 and 3 in any whole number increment that includes an amino acid position having a value greater than 0.5 in the Hydrophilicity profiles of FIG. 5; a peptide region of at least 5 amino adds of FIGS. 2 and 3 in any whole number increment that includes an amino acid position having a value less than 0.5 in the Hydropathicity profile of FIG. 6; a peptide region of at least 5 amino adds of FIGS. 2 and 3 in any whole number increment that includes an amino acid position having a value greater than 0.5 in the Percent Accessible Residues profiles of FIG. 7; a peptide region of at least 5 amino adds of FIGS. 2 and 3 in any whole number increment that includes an amino acid position having a value greater than 0.5 in the Average Flexibility profiles on FIG. 8; and, a peptide region of at least 5 amino adds of FIGS. 2 and 3 in any whole number increment that includes an amino acid position having a value greater than 0.5 in the Beta-turn profile of FIG. 9. Peptide immunogens of the invention can also comprise nucleic adds that encode any of the forgoing.

All immunogens of the invention, peptide or nucleic acid, can be embodied in human unit dose form, or comprised by a composition that includes a pharmaceutical excipient compatible with human physiology.

The secondary structure of 98P4B6 protein variants 1, 2, 5-7, namely the predicted presence and location of alpha helices, extended strands, and random coils, is predicted from the primary amino acid sequence using the HNN—Hierarchical Neural Network method, accessed from the ExPasy molecular biology server. The analysis indicates that 98P4B6 variant 1 is composed of 54.41% alpha helix, 12.33% extended strand, and 33.26% random coil (FIG. 13A). Variant 2 is composed of 17.78% alpha helix, 6.67% extended strand, and 75.56% random coil (FIG. 13B). Variant 5 is composed of 51.55% alpha helix, 13.13% extended strand, and 35.32% random coil (FIG. 13C). Variant 6 is composed of 54.49% alpha helix, 11.84% extended strand, and 33.67% random coil (FIG. 13D). Variant 7 is composed of 48.26% alpha helix, 15.28% extended strand, and 36.46% random coil (FIG. 13E).

Analysis for the potential presence of transmembrane domains in the 98P4B6 variant proteins was carried out using a variety of transmembrane prediction algorithms accessed from the ExPasy molecular biology server. Shown graphically in FIGS. 13F and 13G are the results of analysis of variant 1 depicting the presence and location of 6 transmembrane domains using the TMpred program (FIG. 13F) and 5 transmembrane domains using the TMHMM program (FIG. 13G). Shown graphically in FIGS. 13H and 13I are the results of analysis of variant 2 depicting the presence and location of 1 transmembrane domains using the TMpred program (FIG. 13H) and no transmembrane domains using the TMHMM program (FIG. 13I). Shown graphically in FIGS. 13J and 13K are the results of analysis of variant 5 depicting the presence and location of 6 transmembrane domains using the TMpred program (FIG. 13J) and 4 transmembrane domains using the TMHMM program (FIG. 13K). Shown graphically in FIGS. 13L and 13M are the results of analysis of variant 6 depicting the presence and location of 6 transmembrane domains using the TMpred program (FIG. 13L) and 6 transmembrane domains using the TMHMM program (FIG. 13M). Shown graphically in FIGS. 13N and 13O are the results of analysis of variant 7 depicting the presence and location of 6 transmembrane domains using the TMpred program (FIG. 13N) and 4 transmembrane domains using the TMHMM program (FIG. 13O). The results of each program, namely the amino acids encoding the transmembrane domains are summarized in Table VI.

Example 10 Generation of 98P4B6 Polyclonal Antibodies

Polyclonal antibodies can be raised in a mammal, for example, by one or more injections of an immunizing agent and, if desired, an adjuvant. Typically, the immunizing agent and/or adjuvant will be injected in the mammal by multiple subcutaneous or intraperitoneal injections. In addition to immunizing with a full length 98P4B6 protein variant, computer algorithms are employed in design of immunogens that, based on amino acid sequence analysis contain characteristics of being antigenic and available for recognition by the immune system of the immunized host (see Example 9 entitled “Antigenicity Profiles and Secondary Structure”). Such regions would be predicted to be hydrophilic, flexible, in beta-trn conformations, and be exposed on the surface of the protein (see, e.g., FIG. 5(A-E), FIGS. 6(A & B), FIG. 7(A-E), FIG. 8(A-E), or FIG. 9(A-E) for amino acid profiles that indicate such regions of 98P4B6 protein variants).

For example, recombinant bacterial fusion proteins or peptides containing hydrophilic, flexible, beta-turn regions of 98P4B6 protein variants are used as antigens to generate polyclonal antibodies in New Zealand White rabbits or monoclonal antibodies as described in Example 11. For example, in 98P4B6 variant 1, such regions include, but are not limited to, amino adds 153-165, amino acids 240-260, and amino adds 345-358. In sequence specific for variant 2, such regions include, but are not limited to, amino adds 26-38. In sequence specific for variant 5, such regions include, but are not limited to, amino acids 400-410. In sequence specific for variant 6, such regions include, but are not limited to, amino acids 455-490. In sequence specific for variant 7, such regions include, but are not limited to, amino acids 451-465 and amino adds

472-498. It is useful to conjugate the immunizing agent to a protein known to be immunogenic in the mammal being immunized. Examples of such immunogenic proteins include, but are not limited to, keyhole limpet hemocyanin (KLH), serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. In one embodiment, a peptide encoding amino adds 153-165 of 98P4B6 variant 1 was conjugated to KLH and used to immunize a rabbit. Alternatively the immunizing agent may include all or portions of the 98P4B6 variant proteins, analogs or fusion proteins thereof. For example, the 98P4B6 variant 1 amino acid sequence can be fused using recombinant DNA techniques to any one of a variety of fusion protein partners that are well known in the art, such as glutathione-5-transferase (GST) and HIS tagged fusion proteins. In another embodiment, amino acids 2-204 of 98P4B6 variant 1 was fused to GST using recombinant techniques and the pGEX expression vector, expressed, purified and used to immunize a rabbit. Such fusion proteins are purified from induced bacteria using the appropriate affinity matrix.

Other recombinant bacterial fusion proteins that may be employed include maltose binding protein, LacZ, thioredoxin, NusA, or an immunoglobulin constant region (see the section entitled “Production of 98P4B6 in Prokaryotic Systems” and Current Protocols In Molecular Biology, Volume 2, Unit 16, Frederick M. Ausubul et al. eds., 1995; Linsley, P. S., Brady, W., Urnes, M., Grosmaire, L., Damle, N., and Ledbetter, L. (1991) J. Exp. Med. 174, 561-566).

In addition to bacterial derived fusion proteins, mammalian expressed protein antigens are also used. These antigens are expressed from mammalian expression vectors such as the Tag5 and Fc-fusion vectors (see the section entitled “Production of Recombinant 98P4B6 in Eukaryotic Systems”), and retain post-translational modifications such as glycosylations found in naive protein. In one embodiment, amino acids 324-359 of variant 1, encoding an extracellular loop between transmembrane domains, is cloned into the Tag5 mammalian secretion vector. The recombinant protein is purified by metal chelate chromatography from tissue culture supernatants of 293T cells stably expressing the recombinant vector. The purified Tag5 98P4B6 protein is then used as immunogen.

During the immunization protocol, it is useful to mix or emulsify the antigen in adjuvants that enhance the immune response of the host animal. Examples of adjuvants include, but are not limited to, complete Freund's adjuvant (CFA) and MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).

In a typical protocol, rabbits are initially immunized subcutaneously with up to 200 μg, typically 100-200 μg, of fusion protein or peptide conjugated to KLH mixed in complete Freund's adjuvant (CFA). Rabbits are then injected subcutaneously every two weeks with up to 200 μg, typically 100-200 μg, of the immunogen in incomplete Freund's adjuvant (IFA). Test bleeds are taken approximately 7-10 days following each immunization and used to monitor the titer of the antiserum by ELISA.

To test reactivity and specificity of immune serum, such as the rabbit serum derived from immunization with the Tag5-98P4B6 variant 1 protein, the full-length 98P4B6 variant 1 cDNA is cloned into pcDNA 3.1 myc-his expression vector (Invitrogen, see the Example entitled “Production of Recombinant 98P4B6 in Eukaryotic Systems”. After transfection of the constructs into 293T cells, cell lysates are probed with the anti-98P4B6 serum and with anti-His antibody (Santa Cruz Biotechnologies, Santa Cruz, Calif.) to determine specific reactivity to denatured 98P4B6 protein using the Western blot technique. Detection of 98P486 variant 1 protein expressed in 293T with polyclonal antibodies raised to a GST-fusion protein and peptide is shown in FIGS. 17B and 17C, respectively. In addition, the immune serum is tested by fluorescence microscopy, flow cytometry and immunoprecipitation against 293T and other recombinant 98P4B6-expressing cells to determine specific recognition of native protein. Western blot, immunoprecipitation, fluorescent microscopy, and flow cytometric techniques using cells that endogenously express 98P4B6 are also carried out to test reactivity and specificity.

Anti-serum from rabbits immunized with 98P4B6 variant fusion proteins, such as GST and MBP fusion proteins, are purified by depletion of antibodies reactive to the fusion partner sequence by passage over an affinity column containing the fusion partner either alone or in the context of an irrelevant fusion protein. For example, antiserum derived from a GST-98P4B6 variant 1 fusion protein was first purified by passage over a column of GST protein covalently coupled to AffiGel matrix (BioRad, Hercules, Calif.). The antiserum is then affinity purified by passage over a column composed of a MBP-98P4B6 fusion protein covalently coupled to Affigel matrix. The serum is then further purified by protein G affinity chromatography to isolate the IgG fraction. Sera from other His-tagged antigens and peptide immunized rabbits as well as fusion partner depleted sera are affinity purified by passage over a column matrix composed of the original protein immunogen or free peptide, such as the anti-peptide polyclonal antibody used in FIG. 17C.

Example 11 Generation of 98P4B6 Monoclonal Antibodies (mAbs)

In one embodiment, therapeutic mAbs to 98P4B6 variants comprise those that react with epitopes specific for each variant protein or specific to sequences in common between the variants that would disrupt or modulate the biological function of the 98P4B6 variants, for example those that would disrupt the interaction with ligands and binding partners. Immunogens for generation of such mAbs include those designed to encode or contain the entire 98P4B6 protein variant sequence, regions of the 98P4B6 protein variants predicted to be antigenic from computer analysis of the amino acid sequence (see, e.g., FIG. 5(A-E), FIG. 6(A-E), FIG. 7(A-E), FIG. 8(A-E), or FIG. 9(A-E), and Example 9 entitled “Antigenicity Profiles and Secondary Structure”). Immunogens include peptides, recombinant bacterial proteins, and mammalian expressed Tag 5 proteins and human and murine IgG FC fusion proteins. In addition, cells engineered to express high levels of a respective 98P4B6 variant, such as 293T-98P4B6 variant 1 or 300.19-98P4B6 variant 1murine Pre-B cells, are used to immunize mice.

To generate mAbs to a 98P4B6 variant, mice are first immunized intraperitoneally (IP) with, typically, 10-50 μg of protein immunogen or 10⁷ 98P4B6-expressing cells mixed in complete Freund's adjuvant. Mice are then subsequently immunized IP every 2-4 weeks with, typically, 10-50 μg of protein immunogen or 10⁷ cells mixed in incomplete Freund's adjuvant. Alternatively, MPL-TDM adjuvant is used in immunizations. In addition to the above protein and cell-based immunization strategies, a DNA-based immunization protocol is employed in which a mammalian expression vector encoding a 98P4B6 variant sequence is used to immunize mice by direct injection of the plasmid DNA. For example, amino acids 324-359 is cloned into the Tag5 mammalian secretion vector and the recombinant vector is used as immunogen. In another example the same amino acids are cloned into an Fc-fusion secretion vector in which the 98P4B6 variant 1 sequence is fused at the amino-terminus to an IgK leader sequence and at the carboxyl-terminus to the coding sequence of the human or murine IgG Fc region. This recombinant vector is then used as immunogen. The plasmid immunization protocols are used in combination with purified proteins expressed from the same vector and with cells expressing the respective 98P4B6 variant.

During the immunization protocol, test bleeds are taken 7-10 days following an injection to monitor titer and specificity of the immune response. Once appropriate reactivity and specificity is obtained as determined by ELISA, Western blotting, immunoprecipitation, fluorescence microscopy, and flow cytometric analyses, fusion and hybridoma generation is then carried out with established procedures well known in the art (see, e.g., Harlow and Lane, 1988).

In one embodiment for generating 98P4B6 monoclonal antibodies, a Tag5-98P4B6 variant 1 antigen encoding amino acids 324-359, is expressed and purified from stably transfected 293T cells. Balb C mice are initially immunized intraperitoneally with 25 μg of the Tag5-98P4B6 variant 1 protein mixed in complete Freund's adjuvant. Mice are subsequently immunized every two weeks with 25 μg of the antigen mixed in incomplete Freund's adjuvant for a total of three immunizations. ELISA using the Tag5 antigen determines the titer of serum from immunized mice. Reactivity and specificity of serum to full length 98P4B6 variant 1 protein is monitored by Western blotting, immunoprecipitation and flow cytometry using 293T cells transfected with an expression vector encoding the 98P4B6 variant 1 cDNA (see e.g., the Example entitled “Production of Recombinant 98P4B6 in Eukaryotic Systems” and FIG. 20). Other recombinant 98P4B6 variant 1-expressing cells or cells endogenously expressing 98P4B6 variant 1 are also used. Mice showing the strongest reactivity are rested and given a final injection of Tag5 antigen in PBS and then sacrificed four days later. The spleens of the sacrificed mice are harvested and fused to SPO/2 myeloma cells using standard procedures (Harlow and Lane, 1988). Supernatants from HAT selected growth wells are screened by ELISA, Western blot, immunoprecipitation, fluorescent microscopy, and flow cytometry to identify 98P4B6 specific antibody-producing clones.

To generate monoclonal antibodies that are specific for each 98P4B6 variant protein, immunogens are designed to encode sequences unique for each variant. In one embodiment, a Tag5 antigen encoding the full sequence of 98P486 variant 2 (AA 145) is produced, purified and used as immunogen to derive monoclonal antibodies specific to 98P4B6 variant 2. In another embodiment, an antigenic peptide composed of amino acids 400-410 of 98P4B6 variant 5 is coupled to KLH and used as immunogen. In another embodiment, a GST fusion protein encoding amino acids 455-490 of 98P4B6 of variant 6 is used as immunogen to derive variant 6 specific monoclonal antibodies. In another embodiment, a peptide composed of amino acids 472-498 of variant 7 is coupled to KLH and used as immunogen to generate variant 7 specific monoclonal antibodies. Hybridoma supernatants are then screened on the respective antigen and then further screened on cells expressing the specific variant and cross-screened on cells expressing the other variants to derive variant-specific monoclonal antibodies.

The binding affinity of a 98P4B6 variant monoclonal antibody is determined using standard technologies. Affinity measurements quantify the strength of antibody to epitope binding and are used to help define which 98P4B6 variant monoclonal antibodies preferred for diagnostic or therapeutic use, as appreciated by one of skill in the art. The BIAcore system (Uppsala, Sweden) is a preferred method for determining binding affinity. The BIAcore system uses surface plasmon resonance (SPR, Welford K. 1991, Opt. Quant. Elect. 23:1; Morton and Myszka, 1998, Methods in Enzymology 295: 268) to monitor biomolecular interactions in real time. BIAcore analysis conveniently generates association rate constants, dissociation rate constants, equilibrium dissociation constants, and affinity constants.

Example 12 HLA Class I and Class II Binding Assays

HLA class I and class II binding assays using purified HLA molecules are performed in accordance with disclosed protocols (e.g., PCT publications WO 94/20127 and WO 94/03205; Sidney et al., Current Protocols in Immunology 18.3.1 (1998); Sidney, et al., J. Immunol. 154:247 (1995); Sette, et al., Mol. Immunol. 31:813 (1994)). Briefly, purified MHC molecules (5 to 500 nM) are incubated with various unlabeled peptide inhibitors and 1-10 nM ¹²⁵I-radiolabeled probe peptides as described. Following incubation, MHC-peptide complexes are separated from free peptide by gel filtration and the fraction of peptide bound is determined. Typically, in preliminary experiments, each MHC preparation is titered in the presence of fixed amounts of radiolabeled peptides to determine the concentration of HLA molecules necessary to bind 10-20% of the total radioactivity. All subsequent inhibition and direct binding assays are performed using these HLA concentrations.

Since under these conditions [label]<[HLA] and IC₅₀≧[HLA], the measured IC₅₀ values are reasonable approximations of the true K_(D) values. Peptide inhibitors are typically tested at concentrations ranging from 120 μg/ml to 1.2 ng/ml, and are tested in two to four completely independent experiments. To allow comparison of the data obtained in different experiments, a relative binding figure is calculated for each peptide by dividing the IC₅₀ of a positive control for inhibition by the IC₅₀ for each tested peptide (typically unlabeled versions of the radiolabeled probe peptide). For database purposes, and inter-experiment comparisons, relative binding values are compiled. These values can subsequently be converted back into IC₅₀ nM values by dividing the IC₅₀ nM of the positive controls for inhibition by the relative binding of the peptide of interest. This method of data compilation is accurate and consistent for comparing peptides that have been tested on different days, or with different lots of purified MHC.

Binding assays as outlined above may be used to analyze HLA supermotif and/or HLA motif-bearing peptides (see Table IV).

Example 13 Identification of HLA Supermotif- and Motif-Bearing CTL Candidate Epitopes

HLA vaccine compositions of the invention can include multiple epitopes. The multiple epitopes can comprise multiple HLA supermotifs or motifs to achieve broad population coverage. This example illustrates the identification and confirmation of supermotif- and motif-bearing epitopes for the inclusion in such a vaccine composition. Calculation of population coverage is performed using the strategy described below.

Computer searches and algorithms for identification of supermotif and/or motif-bearing epitopes The searches performed to identify the motif-bearing peptide sequences in the Example entitled “Antigenicity Profiles” and Tables VIII-XXI and XXII-XLIX employ the protein sequence data from the gene product of 98P4B6 set forth in FIGS. 2 and 3, the specific search peptides used to generate the tables are listed in Table VII.

Computer searches for epitopes bearing HLA Class I or Class II supermotifs or motifs are performed as follows. All translated 98P4B6 protein sequences are analyzed using a text string search software program to identify potential peptide sequences containing appropriate HLA binding motifs; such programs are readily produced in accordance with information in the art in view of known motif/supermotif disclosures. Furthermore, such calculations can be made mentally.

Identified A2-, A3-, and DR-supermotif sequences are scored using polynomial algorithms to predict their capacity to bind to specific HLA-Class I or Class II molecules. These polynomial algorithms account for the impact of different amino acids at different positions, and are essentially based on the premise that the overall affinity (or ΔG) of peptide-HLA molecule interactions can be approximated as a linear polynomial function of the type: “ΔG”=a _(1i) ×a _(2i) ×a _(3i) . . . ×a _(ni)

where a_(ji) is a coefficient which represents the effect of the presence of a given amino acid (j) at a given position (i) along the sequence of a peptide of n amino adds. The crucial assumption of this method is that the effects at each position are essentially independent of each other (i.e., independent binding of individual side-chains). When residue j occurs at position i in the peptide, it is assumed to contribute a constant amount j_(i) to the free energy of binding of the peptide irrespective of the sequence of the rest of the peptide.

The method of derivation of specific algorithm coeffidents has been described in Gulukota et al., J. Mol. Biol. 267:1258-126, 1997; (see also Sidney et al., Human Immunol. 45:79-93, 1996; and Southwood et al., J. Immunol. 160:3363-3373, 1998). Briefly, for all i positions, anchor and non-anchor alike, the geometric mean of the average relative binding (ARB) of all peptides carrying j is calculated relative to the remainder of the group, and used as the estimate of j_(i). For Class peptides, if multiple alignments are possible, only the highest scoring alignment is utilized, following an iterative procedure. To calculate an algorithm score of a given peptide in a test set, the ARB values corresponding to the sequence of the peptide are multiplied. If this product exceeds a chosen threshold, the peptide is predicted to bind. Appropriate thresholds are chosen as a function of the degree of stringency of prediction desired.

Selection of HLA-A2 Supertype Cross-Reactive Peptides

Protein sequences from 98P4B6 are scanned utilizing motif identification software, to identify 8-, 9-, 10- and 11-mer sequences containing the HLA-A2-supermotif main anchor specificity. Typically, these sequences are then scored using the protocol described above and the peptides corresponding to the positive-scoring sequences are synthesized and tested for their capacity to bind purified HLA-A*0201 molecules in vitro (HLA-A*0201 is considered a prototype A2 supertype molecule).

These peptides are then tested for the capacity to bind to additional A2-supertype molecules (A*0202, A*0203, A*0206, and A*6802). Peptides that bind to at least three of the five A2-supertype alleles tested are typically deemed A2-supertype cross-reactive binders. Preferred peptides bind at an affinity equal to or less than 500 nM to three or more HLA-A2 supertype molecules.

Selection of HLA-A3 Supermotif-Bearing Epitopes

The 98P4B6 protein sequence(s) scanned above is also examined for the presence of peptides with the HLA-A3-supermotif primary anchors. Peptides corresponding to the HLA A3 supermotif-bearing sequences are then synthesized and tested for binding to HLA-A*0301 and HLA-A*1101 molecules, the molecules encoded by the two most prevalent A3-supertype alleles. The peptides that bind at least one of the two alleles with binding affinities of ≦500 nM, often ≦200 nM, are then tested for binding cross-reactivity to the other common A3-supertype alleles (e.g., A*3101, A*3301, and A*6801) to identify those that can bind at least three of the five HLA-A3-supertype molecules tested.

Selection of HLA-B7 Supermotif Bearing epitopes

The 98P4B6 protein(s) scanned above is also analyzed for the presence of 8-, 9-, 10-, or 1-mer peptides with the HLA-B7-supermotif. Corresponding peptides are synthesized and tested for binding to HLA-B*0702, the molecule encoded by the most common B7-supertype allele (i.e., the prototype B7 supertype allele). Peptides binding B*0702 with IC₅₀ of ≦500 nM are identified using standard methods. These peptides are then tested for binding to other common B7-supertype molecules (e.g., B*3501, B*5101, B*5301, and B*5401). Peptides capable of binding to three or more of the five B7-supertype alleles tested are thereby identified.

Selection of A1 and A24 Motif-Bearing Epitopes

To further increase population coverage, HLA-A1 and -A24 epitopes can also be incorporated into vaccine compositions. An analysis of the 98P4B6-protein can also be performed to identify HLA-A1- and A24-motif-containing sequences.

High affinity and/or cross-reactive binding epitopes that bear other motif and/or supermotifs are identified using analogous methodology.

Example 14 Confirmation of Immunogenicity

Cross-reactive candidate CTL A2-supermotif-bearing peptides that are identified as described herein are selected to confirm in vitro immunogenicity. Confirmation is performed using the following methodology:

Target Cell Lines for Cellular Screening:

The 0.221A2.1 cell line, produced by transferring the HLA-A2.1 gene into the HLA-A, -B, -C null mutant human B-lymphoblastoid cell line 721.221, is used as the peptide-loaded target to measure activity of HLA-A2.1-restricted CTL. This cell line is grown in RPMI-1640 medium supplemented with antibiotics, sodium pyruvate, nonessential amino adds and 10% (vN) heat inactivated FCS. Cells that express an antigen of interest, or transfectants comprising the gene encoding the antigen of interest, can be used as target cells to confirm the ability of peptide-specific CTLs to recognize endogenous antigen.

Primary CTL Induction Cultures:

Generation of Dendritic Cells (DC): PBMCs are thawed in RPMI with 30 μg/ml DNAse, washed twice and resuspended in complete medium (RPMI-1640 plus 5% AB human serum, non-essential amino adds, sodium pyruvate, L-glutamine and penicillin/streptomycin). The monocytes are purified by plating 10×10⁶ PBMC/well in a 6-well plate. After 2 hours at 37° C., the non-adherent cells are removed by gently shaking the plates and aspirating the supernatants. The wells are washed a total of three times with 3 ml RPMI to remove most of the non-adherent and loosely adherent cells. Three ml of complete medium containing 50 ng/ml of GM-CSF and 1,000 U/ml of IL-4 are then added to each well. TNFα is added to the DCs on day 6 at 75 ng/ml and the cells are used for CTL induction cultures on day 7.

Induction of CTL with DC and Peptide: CD8+ T-cells are isolated by positive selection with Dynal immunomagnetic beads (Dynabeads® M-450) and the Detacha-bead® reagent. Typically about 200-250×10⁶ PBMC are processed to obtain 24×10⁶ CD8⁺, T-cells (enough for a 48-well plate culture). Briefly, the PBMCs are thawed in RPMI with 30 μg/ml DNAse, washed once with PBS containing 1% human AB serum and resuspended in PBS/1% AB serum at a concentration of 20×10⁶ cells/ml. The magnetic beads are washed 3 times with PBS/AB serum, added to the cells (140 μl beads/20×10⁶ cells) and incubated for 1 hour at 4° C. with continuous mixing. The beads and cells are washed 4× with PBS/AB serum to remove the nonadherent cells and resuspended at 100×10⁶ cells/ml (based on the original cell number) in PBS/AB serum containing 100 μl/ml Detacha-bead® reagent and 30 μg/ml DNAse. The mixture is incubated for 1 hour at room temperature with continuous mixing. The beads are washed again with PBS/AB/DNAse to collect the CD8+ T-cells. The DC are collected and centrifuged at 1300 rpm for 5-7 minutes, washed once with PBS with 1% BSA, counted and pulsed with 40 μg/ml of peptide at a cell concentration of 1-2×10⁶/ml in the presence of 3 μg/ml β₂-microglobulin for 4 hours at 20° C. The DC are then irradiated (4,200 rads), washed 1 time with medium and counted again.

Setting up induction cultures: 0.25 ml cytokine-generated DC (at 1×10⁵ cells/ml) are co-cultured with 0.25 ml of CD8+ T-cells (at 2×10⁶ cell/ml) in each well of a 48-well plate in the presence of 10 ng/ml of IL-7. Recombinant human IL-10 is added the next day at a final concentration of 10 ng/ml and rhuman IL-2 is added 48 hours later at 10 IU/ml.

Restimulation of the induction cultures with peptide-pulsed adherent cells: Seven and fourteen days after the primary induction, the cells are restimulated with peptide-pulsed adherent cells. The PBMCs are thawed and washed twice with RPMI and DNAse. The cells are resuspended at 5×10⁶ cells/ml and irradiated at ˜4200 rads. The PBMCs are plated at 2×10⁶ in 0.5 ml complete medium per well and incubated for 2 hours at 37° C. The plates are washed twice with RPMI by tapping the plate gently to remove the nonadherent cells and the adherent cells pulsed with 10 μg/ml of peptide in the presence of 3 μg/ml 92 microglobulin in 0.25 ml RPMI/5% AB per well for 2 hours at 37° C. Peptide solution from each well is aspirated and the wells are washed once with RPMI. Most of the media is aspirated from the induction cultures (CD8+ cells) and brought to 0.5 ml with fresh media. The cells are then transferred to the wells containing the peptide-pulsed adherent cells. Twenty four hours later recombinant human IL-10 is added at a final concentration of 10 ng/ml and recombinant human IL-2 is added the next day and again 2-3 days later at 50 IU/ml (Tsai et al., Critical Reviews in Immunology 18(1-2):65-75, 1998). Seven days later, the cultures are assayed for CTL activity in a ⁵¹Cr release assay. In some experiments the cultures are assayed for peptide-specific recognition in the in situ IFNγ ELISA at the time of the second restimulation followed by assay of endogenous recognition 7 days later. After expansion, activity is measured in both assays for a side-by-side comparison.

Measurement of CTL Lytic Activity by ⁵¹Cr Release.

Seven days after the second restimulation, cytotoxicity is determined in a standard (5 hr) ⁵¹Cr release assay by assaying individual wells at a single E:T. Peptide-pulsed targets are prepared by incubating the cells with 10 μg/ml peptide overnight at 37° C.

Adherent target cells are removed from culture flasks with trypsin-EDTA. Target cells are labeled with 200 μCi of ⁵¹Cr sodium chromate (Dupont, Wilmington, Del.) for 1 hour at 37° C. Labeled target cells are resuspended at 10⁶ per ml and diluted 1:10 with K562 cells at a concentration of 3.3×10⁶/ml (an NK-sensitive erythroblastoma cell line used to reduce non-specific lysis). Target cells (100 μl) and effectors (100 μl) are plated in 96 well round-bottom plates and incubated for 5 hours at 37° C. At that time, 100 μl of supernatant are collected from each well and percent lysis is determined according to the formula: [(cpm of the test sample−cpm of the spontaneous ⁵¹Cr release sample)/(cpm of the maximal ⁵¹Cr release sample-−cpm of the spontaneous ⁵¹Cr release sample)]×100.

Maximum and spontaneous release are determined by incubating the labeled targets with 1% Triton X-100 and media alone, respectively. A positive culture is defined as one in which the specific lysis (sample-background) is 10% or higher in the case of individual wells and is 15% or more at the two highest E:T ratios when expanded cultures are assayed.

In situ Measurement of Human IFNγ Production as an Indicator of Peptide-Specific and Endogenous Recognition

Immulon 2 plates are coated with mouse ant-human IFNγ monoclonal antibody (4 μg/ml 0.1M NaHCO₃, pH8.2) overnight at 4° C. The plates are washed with Ca², Mg²⁺-free PBS/0.05% Tween 20 and blocked with PBS/10% FCS for two hours, after which the CTLs (100 μl/well) and targets (100 μ/well) are added to each well, leaving empty wells for the standards and blanks (which received media only). The target cells, either peptide-pulsed or endogenous targets, are used at a concentration of 1×10⁶ cells/ml. The plates are incubated for 48 hours at 37° C. with 5% CO₂.

Recombinant human IFN-gamma is added to the standard wells starting at 400 μg or 1200 μg/100 microliter/well and the plate incubated for two hours at 37° C. The plates are washed and 100 μl of biotinylated mouse anti-human IFN-gamma monoclonal antibody (2 microgram/ml in PBS/3% FCS/0.05% Tween 20) are added and incubated for 2 hours at room temperature. After washing again, 100 microliter HRP-streptavidin (1:4000) are added and the plates incubated for one hour at room temperature. The plates are then washed 6× with wash buffer, 100 microliter/well developing solution (TMB 1:1) are added, and the plates allowed to develop for 5-15 minutes. The reaction is stopped with 50 microliter/well 1M H₃PO₄ and read at OD450. A culture is considered positive if it measured at least 50 μg of IFN-gamma/well above background and is twice the background level of expression.

CTL Expansion.

Those cultures that demonstrate specific lytic activity against peptide-pulsed targets and/or tumor targets are expanded over a two week period with anti-CD3. Briefly, 5×10⁴ CD8+ cells are added to a T25 flask containing the following: 1×10⁶ irradiated (4,200 rad) PBMC (autologous or allogeneic) per ml, 2×10⁵ irradiated (8,000 rad) EBV-transformed cells per ml, and OKT3 (anti-CD3) at 30 ng per ml in RPMI-1640 containing 10% (v/v) human AB serum, non-essential amino acids, sodium pyruvate, 25 μM 2-mercaptoethanol, L-glutamine and penicillin/streptomycin. Recombinant human IL2 is added 24 hours later at a final concentration of 200 IU/ml and every three days thereafter with fresh media at 50 IU/ml. The cells are split if the cell concentration exceeds 1×10⁶/ml and the cultures are assayed between days 13 and 15 at E:T ratios of 30, 10, 3 and 1:1 in the ⁵¹Cr release assay or at 1×10⁶ ml in the in situ IFNγ assay using the same targets as before the expansion.

Cultures are expanded in the absence of anti-CD3, as follows. Those cultures that demonstrate specific lytic activity against peptide and endogenous targets are selected and 5×10⁴ CD8⁺ cells are added to a T25 flask containing the following: 1×10⁶ autologous PBMC per ml which have been peptide-pulsed with 10 μg/ml peptide for two hours at 37° C. and irradiated (4,200 rad); 2×10⁵ irradiated (8,000 rad) EBV-transformed cells per ml RPMI-1640 containing 10% (v/v) human AB serum, non-essential AA, sodium pyruvate, 25 mM 2-ME, L-glutamine and gentamicin.

Immunogenicity of A2 Supermotif-Bearing Peptides

A2-supermotif cross-reactive binding peptides are tested in the cellular assay for the ability to induce peptide-specific CTL in normal individuals. In this analysis, a peptide is typically considered to be an epitope if it induces peptide-specific CTLs in at least individuals, and preferably, also recognizes the endogenously expressed peptide.

Immunogenicity can also be confirmed using PBMCs isolated from patients bearing a tumor that expresses 98P4B6. Briefly, PBMCs are isolated from patients, re-stimulated with peptide-pulsed monocytes and assayed for the ability to recognize peptide-pulsed target cells as well as transfected cells endogenously expressing the antigen.

Evaluation of A*03/A11 Immunogenicity

HLA-A3 supermotif-bearing cross-reactive binding peptides are also evaluated for immunogenicity using methodology analogous for that used to evaluate the immunogenicity of the HLA-A2 supermotif peptides.

Evaluation of B7 Immunogenicity

Immunogenicity screening of the B7-supertype cross-reactive binding peptides identified as set forth herein are confirmed in a manner analogous to the confirmation of A2- and A3-supermotif-bearing peptides.

Peptides bearing other supermotifs/motifs, e.g., HLA-A1, HLA-A24 etc. are also confirmed using similar methodology

Example 15 Implementation of the Extended Supermotif to Improve the Binding Capacity of Native Epitopes by Creating Analogs

HLA motifs and supermotifs (comprising primary and/or secondary residues) are useful in the identification and preparation of highly cross-reactive native peptides, as demonstrated herein. Moreover, the definition of HLA motifs and supermotifs also allows one to engineer highly cross-reactive epitopes by identifying residues within a native peptide sequence which can be analoged to confer upon the peptide certain characteristics, e.g. greater cross-reactivity within the group of HLA molecules that comprise a supertype, and/or greater binding affinity for some or all of those HLA molecules. Examples of analoging peptides to exhibit modulated binding affinity are set forth in this example.

Analoging at Primary Anchor Residues

Peptide engineering strategies are implemented to further increase the cross-reactivity of the epitopes. For example, the main anchors of A2-supermotif-bearing peptides are altered, for example, to introduce a preferred L, I, V, or M at position 2, and I or V at the C-terminus.

To analyze the cross-reactivity of the analog peptides, each engineered analog is initially tested for binding to the prototype A2 supertype allele A*0201, then, if A*0201 binding capacity is maintained, for A2-supertype cross-reactivity.

Alternatively, a peptide is confirmed as binding one or all supertype members and then analoged to modulate binding affinity to any one (or more) of the supertype members to acid population coverage.

The selection of analogs for immunogenicity in a cellular screening analysis is typically further restricted by the capacity of the parent wild type (WT) peptide to bind at least weakly, i.e., bind at an IC₅₀ of 5000 nM or less, to three of more A2 supertype alleles. The rationale for this requirement is that the WT peptides must be present endogenously in sufficient quantity to be biologically relevant. Analoged peptides have been shown to have increased immunogenicity and cross-reactivity by T cells specific for the parent epitope (see, e.g., Parkhurst et al., J. Immunol. 157:2539, 1996; and Pogue et al., Proc. Natl. Acad. Sci. USA 92:8166, 1995).

In the cellular screening of these peptide analogs, it is important to confirm that analog-specific CTLs are also able to recognize the wild-type peptide and, when possible, target cells that endogenously express the epitope.

Analoging of HLA-A3 and B7-Supermotif-Bearing Peptides

Analogs of HLA-A3 supermotif-bearing epitopes are generated using strategies similar to those employed in analoging HLA-A2 supermotif-bearing peptides. For example, peptides binding to 3/5 of the A3-supertype molecules are engineered at primary anchor residues to possess a preferred residue (V, S, M, or A) at position 2.

The analog peptides are then tested for the ability to bind A*03 and A*11 (prototype A3 supertype alleles). Those peptides that demonstrate ≦500 nM binding capacity are then confirmed as having A3-supertype cross-reactivity.

Similarly to the A2- and A3-motif bearing peptides, peptides binding 3 or more B7-supertype alleles can be improved, where possible, to achieve increased cross-reactive binding or greater binding affinity or binding half life. B7 supermotif-bearing peptides are, for example, engineered to possess a preferred residue (V, I, L, or F) at the C-terminal primary anchor position, as demonstrated by Sidney et al. (J. Immunol. 157:3480-3490, 1996).

Analoging at primary anchor residues of other motif and/or supermotif-bearing epitopes is performed in a like manner.

The analog peptides are then be confirmed for immunogenicity, typically in a cellular screening assay. Again, it is generally important to demonstrate that analog-specific CTLs are also able to recognize the wild-type peptide and, when possible, targets that endogenously express the epitope.

Analoging at Secondary Anchor Residues

Moreover, HLA supermotifs are of value in engineering highly cross-reactive peptides and/or peptides that bind HLA molecules with increased affinity by identifying particular residues at secondary anchor positions that are associated with such properties. For example, the binding capacity of a B7 supermotif-bearing peptide with an F residue at position 1 is analyzed. The peptide is then analoged to, for example, substitute L for F at position 1. The analoged peptide is evaluated for increased binding affinity, binding half life and/or increased cross-reactivity. Such a procedure identifies analoged peptides with enhanced properties.

Engineered analogs with sufficiently improved binding capacity or cross-reactivity can also be tested for immunogenicity in HLA-B7-transgenic mice, following for example, IFA immunization or lipopeptide immunization. Analoged peptides are additionally tested for the ability to stimulate a recall response using PBMC from patients with 98P4B6-expressing tumors.

Other Analoging Strategies

Another form of peptide analoging, unrelated to anchor positions, involves the substitution of a cysteine with α-amino butyric acid. Due to its chemical nature, cysteine has the propensity to form disulfide bridges and sufficiently alter the peptide structurally so as to reduce binding capacity. Substitution of α-amino butyric acid for cysteine not only alleviates this problem, but has been shown to improve binding and crossbinding capabilities in some instances (see, e.g., the review by Sette et al., In: Persistent Viral Infections, Eds. R. Ahmed and I. Chen, John Wiley & Sons, England, 1999).

Thus, by the use of single amino acid substitutions, the binding properties and/or cross-reactivity of peptide ligands for HLA supertype molecules can be modulated.

Example 16 Identification and Confirmation of 98P4B6-Derived Sequences with HLA-DR Binding Motifs

Peptide epitopes bearing an HLA class II supermotif or motif are identified and confirmed as outlined below using methodology similar to that described for HLA Class I peptides.

Selection of HLA-DR-Supermotif-Bearing Epitopes.

To identify 98P4B6-derived, HLA class II HTL epitopes, a 98P4B6 antigen is analyzed for the presence of sequences bearing an HLA-DR-motif or supermotif. Specifically, 15-mer sequences are selected comprising a DR-supermotif, comprising a 9-mer core, and three-residue N- and C-terminal flanking regions (15 amino acids total).

Protocols for predicting peptide binding to DR molecules have been developed (Southwood et al., J. Immunol. 160:3363-3373, 1998). These protocols, specific for individual DR molecules, allow the scoring, and ranking, of 9-mer core regions. Each protocol not only scores peptide sequences for the presence of DR-supermotif primary anchors (i.e., at position 1 and position 6) within a 9-mer core, but additionally evaluates sequences for the presence of secondary anchors. Using allele-specific selection tables (see, e.g., Southwood et al., ibid.), it has been found that these protocols efficiently select peptide sequences with a high probability of binding a particular DR molecule. Additionally, it has been found that performing these protocols in tandem, specifically those for DR1, DR4w4, and DR7, can efficiently select DR cross-reactive peptides.

The 98P4B6-derived peptides identified above are tested for their binding capacity for various common HLA-DR molecules. All peptides are initially tested for binding to the DR molecules in the primary panel: DR1, DR4w4, and DR7. Peptides binding at least two of these three DR molecules are then tested for binding to DR2w2 β1, DR2w2 β2, DR6w19, and DR9 molecules in secondary assays. Finally, peptides binding at least two of the four secondary panel DR molecules, and thus cumulatively at least four of seven different DR molecules, are screened for binding to DR4w15, DR5w11, and DR8w2 molecules in tertiary assays. Peptides binding at least seven of the ten DR molecules comprising the primary, secondary, and tertiary screening assays are considered cross-reactive DR binders. 98P4B6-derived peptides found to bind common HLA-DR alleles are of particular interest.

Selection of DR3 Motif Peptides

Because HLA-DR3 is an allele that is prevalent in Caucasian, Black, and Hispanic populations, DR3 binding capacity is a relevant criterion in the selection of HTL epitopes. Thus, peptides shown to be candidates may also be assayed for their DR3 binding capacity. However, in view of the binding specificity of the DR3 motif, peptides binding only to DR3 can also be considered as candidates for inclusion in a vaccine formulation.

To efficiently identify peptides that bind DR3, target 98P4B6 antigens are analyzed for sequences carrying one of the two DR3-specific binding motifs reported by Geluk et al. (J. Immunol. 152:5742-5748, 1994). The corresponding peptides are then synthesized and confirmed as having the ability to bind DR3 with an affinity of 1 μM or better, i.e., less than 1 μM. Peptides are found that meet this binding criterion and qualify as HLA class II high affinity binders.

DR3 binding epitopes identified in this manner are included in vaccine compositions with DR supermotif-bearing peptide epitopes.

Similarly to the case of HLA class I motif-bearing peptides, the class II motif-bearing peptides are analoged to improve affinity or cross-reactivity. For example, aspartic acid at position 4 of the 9-mer core sequence is an optimal residue for DR3 binding, and substitution for that residue often improves DR3 binding.

Example 17 Immunogenicity of 98P4B6-Derived HTL Epitomes

This example determines immunogenic DR supermotif- and DR3 motif-bearing epitopes among those identified using the methodology set forth herein.

Immunogenicity of HTL epitopes are confirmed in a manner analogous to the determination of immunogenicity of CTL epitopes, by assessing the ability to stimulate HTL responses and/or by using appropriate transgenic mouse models. Immunogenicity is determined by screening for: 1.) in vitro primary induction using normal PBMC or 2.) recall responses from patients who have 98P4B6-expressing tumors.

Example 18 Calculation of Phenotypic Frequencies of HLA-Supertypes in Various Ethnic Backgrounds to Determine Breadth of Population Coverage

This example illustrates the assessment of the breadth of population coverage of a vaccine composition comprised of multiple epitopes comprising multiple supermotifs and/or motifs.

In order to analyze population coverage, gene frequencies of HLA alleles are determined. Gene frequencies for each HLA allele are calculated from antigen or allele frequencies utilizing the binomial distribution formulae gf=1−(SQRT(1−af)) (see, e.g., Sidney et al., Human Immunol. 45:79-93, 1996). To obtain overall phenotypic frequencies, cumulative gene frequencies are calculated, and the cumulative antigen frequencies derived by the use of the inverse formula [af−1−(1−Cgf)²].

Where frequency data is not available at the level of DNA typing, correspondence to the serologically defined antigen frequencies is assumed. To obtain total potential supertype population coverage no linkage disequilibrium is assumed, and only alleles confirmed to belong to each of the supertypes are included (minimal estimates). Estimates of total potential coverage achieved by inter-loci combinations are made by adding to the A coverage the proportion of the non-A covered population that could be expected to be covered by the B alleles considered (e.g., total=A+B*(1−A)). Confirmed members of the A3-like supertype are A3, A11, A31, A*3301, and A*6801. Although the A3-like supertype may also include A34, A66, and A*7401, these alleles were not included in overall frequency calculations. Likewise, confirmed members of the A2-like supertype family are A*0201, A*0202, A*0203, A*0204, A*0205, A*0206, A*0207, A*6802, and A*6901. Finally, the B7-like supertype-confirmed alleles are: B7, B*3501-03, B51, B*5301, B*5401, B*5501-2, B*5601, B*6701, and B*7801 (potentially also B*1401, B*3504-06, B*4201, and B*5602).

Population coverage achieved by combining the A2-, A3- and B7-supertypes is approximately 86% in five major ethnic groups. Coverage may be extended by including peptides bearing the A1 and A24 motifs. On average, A1 is present in 12% and A24 in 29% of the population across five different major ethnic groups (Caucasian, North American Black, Chinese, Japanese, and Hispanic). Together, these alleles are represented with an average frequency of 39% in these same ethnic populations. The total coverage across the major ethnicities when A1 and A24 are combined with the coverage of the A2-, A3- and B7-supertype alleles is >95%, see, e.g., Table IV (G). An analogous approach can be used to estimate population coverage achieved with combinations of class II motif-bearing epitopes.

Immunogenicity studies in humans (e.g., Bertoni et al., J. Clin. Invest. 100:503, 1997; Doolan et al., Immunity 7:97, 1997; and Threlkeld et al., J. Immunol. 159:1648, 1997) have shown that highly cross-reactive binding peptides are almost always recognized as epitopes. The use of highly cross-reactive binding peptides is an important selection criterion in identifying candidate epitopes for inclusion in a vaccine that is immunogenic in a diverse population.

With a sufficient number of epitopes (as disclosed herein and from the art), an average population coverage is predicted to be greater than 95% in each of five major ethnic populations. The game theory Monte Carlo simulation analysis, which is known in the art (see e.g., Osborne, M. J. and Rubinstein, A. “A course in game theory” MIT Press, 1994), can be used to estimate what percentage of the individuals in a population comprised of the Caucasian, North American Black, Japanese, Chinese, and Hispanic ethnic groups would recognize the vaccine epitopes described herein. A preferred percentage is 90%. A more preferred percentage is 95%.

Example 19 CTL Recognition of Endogenously Processed Antigens After Priming

This example confirms that CTL induced by native or analoged peptide epitopes identified and selected as described herein recognize endogenously synthesized, i.e., native antigens.

Effector cells isolated from transgenic mice that are immunized with peptide epitopes, for example HLA-A2 supermotif-bearing epitopes, are re-stimulated in vitro using peptide-coated stimulator cells. Six days later, effector cells are assayed for cytotoxicity and the cell lines that contain peptide-specific cytotoxic activity are further re-stimulated. An additional six days later, these cell lines are tested for cytotoxic activity on ⁵¹Cr labeled Jurkat-A2.1/K^(b) target cells in the absence or presence of peptide, and also tested on ⁵¹Cr labeled target cells bearing the endogenously synthesized antigen, i.e. cells that are stably transfected with 98P4B6 expression vectors.

The results demonstrate that CTL lines obtained from animals primed with peptide epitope recognize endogenously synthesized 98P4B6 antigen. The choice of transgenic mouse model to be used for such an analysis depends upon the epitope(s) that are being evaluated. In addition to HLA-A*0201/K^(b) transgenic mice, several other transgenic mouse models including mice with human A11, which may also be used to evaluate A3 epitopes, and B7 alleles have been characterized and others (e.g., transgenic mice for HLA-A1 and A24) are being developed. HLA-DR1 and HLA-DR3 mouse models have also been developed, which may be used to evaluate HTL epitopes.

Example 20 Activity of CTL-HTL Conjugated Epitopes in Transgenic Mice

This example illustrates the induction of CTLs and HTLs in transgenic mice, by use of a 98P4B6-derived CTL and HTL peptide vaccine compositions. The vaccine composition used herein comprise peptides to be administered to a patient with a 98P4B6-expressing tumor. The peptide composition can comprise multiple CTL and/or HTL epitopes. The epitopes are identified using methodology as described herein. This example also illustrates that enhanced immunogenicity can be achieved by inclusion of one or more HTL epitopes in a CTL vaccine composition; such a peptide composition can comprise an HTL epitope conjugated to a CTL epitope. The CTL epitope can be one that binds to multiple HLA family members at an affinity of 500 nM or less, or analogs of that epitope. The peptides may be lipidated, if desired.

Immunization procedures: Immunization of transgenic mice is performed as described (Alexander et al., J. Immunol. 159:4753-4761, 1997). For example, A2/K^(b) mice, which are transgenic for the human HLA A2.1 allele and are used to confirm the immunogenicity of HLA-A*0201 motif- or HLA-A2 supermotif-bearing epitopes, and are primed subcutaneously (base of the tail) with a 0.1 ml of peptide in Incomplete Freund's Adjuvant, or if the peptide composition is a lipidated CTL/HTL conjugate, in DMSO/saline, or if the peptide composition is a polypeptide, in PBS or Incomplete Freund's Adjuvant. Seven days after priming, splenocytes obtained from these animals are restimulated with syngenic irradiated LPS-activated lymphoblasts coated with peptide.

Cell lines: Target cells for peptide-specific cytotoxicity assays are Jurkat cells transfected with the HLA-A2.1/K^(b) chimeric gene (e.g., Vitiello et al., J. Exp. Med. 173:1007, 1991)

In vitro CTL activation: One week after priming, spleen cells (30×10⁶ cells/flask) are co-cultured at 37° C. with syngeneic, irradiated (3000 rads), peptide coated lymphoblasts (10×10⁶ cells/flask) in 10 ml of culture medium/T25 flask. After six days, effector cells are harvested and assayed for cytotoxic activity.

Assay for cytotoxic activity: Target cells (1.0 to 1.5×10⁶) are incubated at 37° C. in the presence of 200 μl of ⁵¹Cr. After 60 minutes, cells are washed three times and resuspended in R10 medium. Peptide is added where required at a concentration of 1 μg/ml. For the assay, 10⁴ ⁵¹Cr-labeled target cells are added to different concentrations of effector cells (final volume of 200 μl) in U-bottom 96-well plates. After a six hour incubation period at 37° C., a 0.1 ml aliquot of supernatant is removed from each well and radioactivity is determined in a Micromedic automatic gamma counter. The percent specific lysis is determined by the formula: percent specific release=100× (experimental release−spontaneous release)/(maximum release−spontaneous release). To facilitate comparison between separate CTL assays run under the same conditions, % ⁵¹Cr release data is expressed as lytic units/10⁶ cells. One lytic unit is arbitrarily defined as the number of effector cells required to achieve 30% lysis of 10,000 target cells in a six hour ⁵¹Cr release assay. To obtain specific lytic units/10⁶, the lytic units/10⁶ obtained in the absence of peptide is subtracted from the lytic units/10⁶ obtained in the presence of peptide. For example, if 30% ⁵¹Cr release is obtained at the effector (E):target (T) ratio of 50:1 (i.e., 5×10⁵ effector cells for 10,000 targets) in the absence of peptide and 5:1 (i.e., 5×10⁴ effector cells for 10,000 targets) in the presence of peptide, the specific lytic units would be: [(1/50,000)−(1/500,000)]×10⁶=18 LU.

The results are analyzed to assess the magnitude of the CTL responses of animals injected with the immunogenic CTL/HTL conjugate vaccine preparation and are compared to the magnitude of the CTL response achieved using, for example, CTL epitopes as outlined above in the Example entitled “Confirmation of Immunogenicity.” Analyses similar to this may be performed to confirm the immunogenicity of peptide conjugates containing multiple CTL epitopes and/or multiple HTL epitopes. In accordance with these procedures, it is found that a CTL response is induced, and concomitantly that an HTL response is induced upon administration of such compositions.

Example 21 Selection of CTL and HTL Epitopes for Inclusion in a 98P4B6-Specific Vaccine

This example illustrates a procedure for selecting peptide epitopes for vaccine compositions of the invention. The peptides in the composition can be in the form of a nucleic acid sequence, either single or one or more sequences (i.e., minigene) that encodes peptide(s), or can be single and/or polyepitopic peptides.

The following principles are utilized when selecting a plurality of epitopes for inclusion in a vaccine composition. Each of the following principles is balanced in order to make the selection.

Epitopes are selected which, upon administration, mimic immune responses that are correlated with 98P4B6 clearance. The number of epitopes used depends on observations of patients who spontaneously clear 98P4B6. For example, if it has been observed that patients who spontaneously clear 98P4B6-expressing cells generate an immune response to at least three (3) epitopes from 98P4B6 antigen, then at least three epitopes should be included for HLA class-1. A similar rationale is used to determine HLA class II epitopes.

Epitopes are often selected that have a binding affinity of an IC₅₀ of 500 nM or less for an HLA class I molecule, or for class II, an IC₅₀ of 1000 nM or less; or HLA Class I peptides with high binding scores from the BIMAS web site.

In order to achieve broad coverage of the vaccine through out a diverse population, sufficient supermotif bearing peptides, or a sufficient array of allele-specific motif bearing peptides, are selected to give broad population coverage. In one embodiment, epitopes are selected to provide at least 80% population coverage. A Monte Carlo analysis, a statistical evaluation known in the art, can be employed to assess breadth, or redundancy, of population coverage.

When creating polyepitopic compositions, or a minigene that encodes same, it is typically desirable to generate the smallest peptide possible that encompasses the epitopes of interest. The principles employed are similar, if not the same, as those employed when selecting a peptide comprising nested epitopes. For example, a protein sequence for the vaccine composition is selected because it has maximal number of epitopes contained within the sequence, i.e., it has a high concentration of epitopes. Epitopes may be nested or overlapping (i.e., frame shifted relative to one another). For example, with overlapping epitopes, two 9-mer epitopes and one 10-mer epitope can be present in a 10 amino acid peptide. Each epitope can be exposed and bound by an HLA molecule upon administration of such a peptide. A multi-epitopic, peptide can be generated synthetically, recombinantly, or via cleavage from the native source. Alternatively, an analog can be made of this native sequence, whereby one or more of the epitopes comprise substitutions that alter the cross-reactivity and/or binding affinity properties of the polyepitopic peptide. Such a vaccine composition is administered for therapeutic or prophylactic purposes. This embodiment provides for the possibility that an as yet undiscovered aspect of immune system processing will apply to the native nested sequence and thereby facilitate the production of therapeutic or prophylactic immune response-inducing vaccine compositions. Additionally such an embodiment provides for the possibility of motif-bearing epitopes for an HLA makeup that is presently unknown. Furthermore, this embodiment (absent the creating of any analogs) directs the immune response to multiple peptide sequences that are actually present in 98P4B6, thus avoiding the need to evaluate any junctional epitopes. Lastly, the embodiment provides an economy of scale when producing nucleic acid vaccine compositions. Related to this embodiment, computer programs can be derived in accordance with principles in the art, which identify in a target sequence, the greatest number of epitopes per sequence length.

A vaccine composition comprised of selected peptides, when administered, is safe, efficacious, and elicits an immune response similar in magnitude to an immune response that controls or clears cells that bear or overexpress 98P4B6.

Example 22 Construction of “Minigene” Multi-Epitope DNA Plasmids

This example discusses the construction of a minigene expression plasmid. Minigene plasmids may, of course, contain various configurations of B cell, CTL and/or HTL epitopes or epitope analogs as described herein.

A minigene expression plasmid typically includes multiple CTL and HTL peptide epitopes. In the present example, HLA-A2, -A3, -B7 supermotif-bearing peptide epitopes and HLA-A1 and -A24 motif-bearing peptide epitopes are used in conjunction with DR supermotif-bearing epitopes and/or DR3 epitopes. HLA class I supermotif or motif-bearing peptide epitopes derived 98P4B6, are selected such that multiple supermotifs/motifs are represented to ensure broad population coverage. Similarly, HLA class II epitopes are selected from 98P4B6 to provide broad population coverage, i.e. both HLA DR-1-4-7 supermotif-bearing epitopes and HLA DR-3 motif-bearing epitopes are selected for inclusion in the minigene construct. The selected CTL and HTL epitopes are then incorporated into a minigene for expression in an expression vector.

Such a construct may additionally include sequences that direct the HTL epitopes to the endoplasmic reticulum. For example, the li protein may be fused to one or more HTL epitopes as described in the art, wherein the CLIP sequence of the li protein is removed and replaced with an HLA class II epitope sequence so that HLA class II epitope is directed to the endoplasmic reticulum, where the epitope binds to an HLA class II molecules.

This example illustrates the methods to be used for construction of a minigene-bearing expression plasmid. Other expression vectors that may be used for minigene compositions are available and known to those of skill in the art.

The minigene DNA plasmid of this example contains a consensus Kozak sequence and a consensus murine kappa Ig-light chain signal sequence followed by CTL and/or HTL epitopes selected in accordance with principles disclosed herein. The sequence encodes an open reading frame fused to the Myc and His antibody epitope tag coded for by the pcDNA 3.1 Myc-His vector.

Overlapping oligonucleotides that can, for example, average about 70 nucleotides in length with 15 nucleotide overlaps, are synthesized and HPLC-purified. The oligonucleotides encode the selected peptide epitopes as well as appropriate linker nucleotides, Kozak sequence, and signal sequence. The final multiepitope minigene is assembled by extending the overlapping oligonucleotides in three sets of reactions using PCR. A Perkin/Elmer 9600 PCR machine is used and a total of 30 cycles are performed using the following conditions: 95° C. for 15 sec, annealing temperature (50 below the lowest calculated Tm of each primer pair) for 30 sec, and 72° C. for 1 min.

For example, a minigene is prepared as follows. For a first PCR reaction, 5 μg of each of two oligonucleotides are annealed and extended: In an example using eight oligonucleotides, i.e., four pairs of primers, oligonucleotides 1+2, 3+4, 5+6, and 7+8 are combined in 100 μl reactions containing Pfu polymerase buffer (1x=10 mM KCL, 10 mM (NH₄)₂SO₄, 20 mM Tris-chloride, pH 8.75, 2 mM MgSO₄, 0.1% Triton X-100, 100 μg/ml BSA), 0.25 mM each dNTP, and 2.5 U of Pfu polymerase. The full-length dimer products are gel-purified, and two reactions containing the product of 1+2 and 3+4, and the product of 5+6 and 7+8 are mixed, annealed, and extended for 10 cycles. Half of the two reactions are then mixed, and 5 cycles of annealing and extension carried out before flanking primers are added to amplify the full length product. The full-length product is gel-purified and cloned into pCR-blunt (Invitrogen) and individual clones are screened by sequencing.

Example 23 The Plasmid Construct and the Degree to which it Induces Immunogenicity

The degree to which a plasmid construct, for example a plasmid constructed in accordance with the previous Example, is able to induce immunogenicity is confirmed in vitro by determining epitope presentation by APC following transduction or transfection of the APC with an epitope-expressing nucleic acid construct. Such a study determines “antigenicty” and allows the use of human APC. The assay determines the ability of the epitope to be presented by the APC in a context that is recognized by a T cell by quantifying the density of epitope-HLA class I complexes on the cell surface. Quantitation can be performed by directly measuring the amount of peptide eluted from the APC (see, e.g., Sijts et al., J. Immunol. 156:683-692, 1996; Demotz et al., Nature 342:682-684, 1989); or the number of peptide-HLA class I complexes can be estimated by measuring the amount of lysis or lymphokine release induced by diseased or transfected target cells, and then determining the concentration of peptide necessary to obtain equivalent levels of lysis or lymphokine release (see, e.g., Kageyama et al., J. Immunol. 154:567-576, 1995).

Alternatively, immunogenicity is confirmed through in vivo injections into mice and subsequent in vitro assessment of CTL and HTL activity, which are analyzed using cytotoxicity and proliferation assays, respectively, as detailed e.g., in Alexander et al., Immunity 1:751-761, 1994.

For example, to confirm the capacity of a DNA minigene construct containing at least one HLA-A2 supermotif peptide to induce CTLs in vivo, HLA-A2.1/K^(b) transgenic mice, for example, are immunized intramuscularly with 100 μg of naked cDNA. As a means of comparing the level of CTLs induced by cDNA immunization, a control group of animals is also immunized with an actual peptide composition that comprises multiple epitopes synthesized as a single polypeptide as they would be encoded by the minigene.

Splenocytes from immunized animals are stimulated twice with each of the respective compositions (peptide epitopes encoded in the minigene or the polyepitopic peptide), then assayed for peptide-specific cytotoxic activity in a ⁵¹Cr release assay. The results indicate the magnitude of the CTL response directed against the A2-restricted epitope, thus indicating the in vivo immunogenicity of the minigene vaccine and polyepitopic vaccine.

It is, therefore, found that the minigene elicits immune responses directed toward the HLA-A2 supermotif peptide epitopes as does the polyepitopic peptide vaccine. A similar analysis is also performed using other HLA-A3 and HLA-B7 transgenic mouse models to assess CTL induction by HLA-A3 and HLA-B7 motif or supermotif epitopes, whereby it is also found that the minigene elicits appropriate immune responses directed toward the provided epitopes.

To confirm the capacity of a class II epitope-encoding minigene to induce HTLs in vivo, DR transgenic mice, or for those epitopes that cross react with the appropriate mouse MHC molecule, I-A^(b)-restricted mice, for example, are immunized intramuscularly with 100 μg of plasmid DNA. As a means of comparing the level of HTLs induced by DNA immunization, a group of control animals is also immunized with an actual peptide composition emulsified in complete Freund's adjuvant. CD4+ T cells, i.e. HTLs, are purified from splenocytes of immunized animals and stimulated with each of the respective compositions (peptides encoded in the minigene). The HTL response is measured using a ³H-thymidine incorporation proliferation assay, (see, e.g., Alexander et al. Immunity 1:751-761, 1994). The results indicate the magnitude of the HTL response, thus demonstrating the in vivo immunogenicity of the minigene.

DNA minigenes, constructed as described in the previous Example, can also be confirmed as a vaccine in combination with a boosting agent using a prime boost protocol. The boosting agent can consist of recombinant protein (e.g., Barnett et al., Aids Res. and Human Retroviruses 14, Supplement 3:S299-S309, 1998) or recombinant vaccinia, for example, expressing a minigene or DNA encoding the complete protein of interest (see, e.g., Hanke et al., Vaccine 16:439-445, 1998; Sedegah et al., Proc. Natl. Acad. Sci. USA 95:7648-53, 1998; Hanke and McMichael, Immunol. Letters 66:177-181, 1999; and Robinson et al., Nature Med. 5:526-34, 1999).

For example, the efficacy of the DNA minigene used in a prime boost protocol is initially evaluated in transgenic mice. In this example, A2.1/K^(b) transgenic mice are immunized IM with 100 μg of a DNA minigene encoding the immunogenic peptides including at least one HLA-A2 supermotif-bearing peptide. After an incubation period (ranging from 3-9 weeks), the mice are boosted IP with 10⁷ pfu/mouse of a recombinant vaccinia virus expressing the same sequence encoded by the DNA minigene. Control mice are immunized with 100 μg of DNA or recombinant vaccinia without the minigene sequence, or with DNA encoding the minigene, but without the vaccinia boost. After an additional incubation period of two weeks, splenocytes from the mice are immediately assayed for peptide-specific activity in an ELISPOT assay. Additionally, splenocytes are stimulated in vitro with the A2-restricted peptide epitopes encoded in the minigene and recombinant vaccinia, then assayed for peptide-specific activity in an alpha, beta and/or gamma IFN ELISA.

It is found that the minigene utilized in a prime-boost protocol elicits greater immune responses toward the HLA-A2 supermotif peptides than with DNA alone. Such an analysis can also be performed using HLA-A11 or HLA-B7 transgenic mouse models to assess CTL induction by HLA-A3 or HLA-B7 motif or supermotif epitopes. The use of prime boost protocols in humans is described below in the Example entitled “Induction of CTL Responses Using a Prime Boost Protocol.”

Example 24 Peptide Compositions for Prophylactic Uses

Vaccine compositions of the present invention can be used to prevent 98P4B6 expression in persons who are at risk for tumors that bear this antigen. For example, a polyepitopic peptide epitope composition (or a nucleic acid comprising the same) containing multiple CTL and HTL epitopes such as those selected in the above Examples, which are also selected to target greater than 80% of the population, is administered to individuals at risk for a 98P4B6-associated tumor.

For example, a peptide-based composition is provided as a single polypeptide that encompasses multiple epitopes. The vaccine is typically administered in a physiological solution that comprises an adjuvant, such as Incomplete Freunds Adjuvant. The dose of peptide for the initial immunization is from about 1 to about 50,000 μg, generally 100-5,000 μg, for a 70 kg patient. The initial administration of vaccine is followed by booster dosages at 4 weeks followed by evaluation of the magnitude of the immune response in the patient, by techniques that determine the presence of epitope-specific CTL populations in a PBMC sample. Additional booster doses are administered as required. The composition is found to be both safe and efficacious as a prophylaxis against 98P4B6-associated disease.

Alternatively, a composition typically comprising transfecting agents is used for the administration of a nucleic acid-based vaccine in accordance with methodologies known in the art and disclosed herein.

Example 25 Polyepitopic Vaccine Compositions Derived from Native 98P4B6 Sequences

A native 98P4B6 polyprotein sequence is analyzed, preferably using computer algorithms defined for each class I and/or class II supermotif or motif, to identify “relatively short” regions of the polyprotein that comprise multiple epitopes. The “relatively short” regions are preferably less in length than an entire native antigen. This relatively short sequence that contains multiple distinct or overlapping, “nested” epitopes can be used to generate a minigene construct. The construct is engineered to express the peptide, which corresponds to the native protein sequence. The “relatively short” peptide is generally less than 250 amino adds in length, often less than 100 amino adds in length, preferably less than 75 amino acids in length, and more preferably less than 50 amino acids in length. The protein sequence of the vaccine composition is selected because it has maximal number of epitopes contained within the sequence, i.e., it has a high concentration of epitopes. As noted herein, epitope motifs may be nested or overlapping (i.e., frame shifted relative to one another). For example, with overlapping epitopes, two 9-mer epitopes and one 10-mer epitope can be present in a 10 amino acid peptide. Such a vaccine composition is administered for therapeutic or prophylactic purposes.

The vaccine composition will include, for example, multiple CTL epitopes from 98P4B6 antigen and at least one HTL epitope. This polyepitopic native sequence is administered either as a peptide or as a nucleic acid sequence which encodes the peptide. Alternatively, an analog can be made of this native sequence, whereby one or more of the epitopes comprise substitutions that alter the cross-reactivity and/or binding affinity properties of the polyepitopic peptide.

The embodiment of this example provides for the possibility that an as yet undiscovered aspect of immune system processing will apply to the native nested sequence and thereby facilitate the production of therapeutic or prophylactic immune response-including vaccine compositions. Additionally, such an embodiment provides for the possibility of motif-bearing epitopes for an HLA makeup(s) that is presently unknown. Furthermore, this embodiment (excluding an analoged embodiment) directs the immune response to multiple peptide sequences that are actually present in native 98P4B6, thus avoiding the need to evaluate any junctional epitopes. Lastly, the embodiment provides an economy of scale when producing peptide or nucleic acid vaccine compositions.

Related to this embodiment, computer programs are available in the art which can be used to identify in a target sequence, the greatest number of epitopes per sequence length.

Example 26 Polyepitopic Vaccine Compositions from Multiple Antigens

The 98P4B6 peptide epitopes of the present invention are used in conjunction with epitopes from other target tumor-associated antigens, to create a vaccine composition that is useful for the prevention or treatment of cancer that expresses 98P4B6 and such other antigens. For example, a vaccine composition can be provided as a single polypeptide that incorporates multiple epitopes from 98P4B6 as well as tumor-associated antigens that are often expressed with a target cancer associated with 98P4B6 expression, or can be administered as a composition comprising a cocktail of one or more discrete epitopes. Alternatively, the vaccine can be administered as a minigene construct or as dendritic cells which have been loaded with the peptide epitopes in vitro.

Example 27 Use of Peptides to Evaluate an Immune Response

Peptides of the invention may be used to analyze an immune response for the presence of specific antibodies, CTL or HTL directed to 98P4B6. Such an analysis can be performed in a manner described by Ogg et al., Science 279:2103-2106, 1998. In this Example, peptides in accordance with the invention are used as a reagent for diagnostic or prognostic purposes, not as an immunogen.

In this example highly sensitive human leukocyte antigen tetrameric complexes (“tetramers”) are used for a cross-sectional analysis of, for example, 98P4B6 HLA-A*0201-specific CTL frequencies from HLA A*0201-positive individuals at different stages of disease or following immunization comprising a 98P4B6 peptide containing an A*0201 motif. Tetrameric complexes are synthesized as described (Musey et al., N. Engl. J. Med. 337:1267, 1997). Briefly, purified HLA heavy chain (A*0201 in this example) and β2-microglobulin are synthesized by means of a prokaryotic expression system. The heavy chain is modified by deletion of the transmembrane-cytosolic tail and COOH-terminal addition of a sequence containing a BirA enzymatic biotinylation site. The heavy chain, P2-microglobulin, and peptide are refolded by dilution. The 45-kD refolded product is isolated by fast protein liquid chromatography and then biotinylated by BirA in the presence of biotin (Sigma, St. Louis, Mo.), adenosine 5′ triphosphate and magnesium. Streptavidin-phycoerythrin conjugate is added in a 1:4 molar ratio, and the tetrameric product is concentrated to 1 mg/ml. The resulting product is referred to as tetramer-phycoerythrin.

For the analysis of patient blood samples, approximately one million PBMCs are centrifuged at 300 g for 5 minutes and resuspended in 50 μl of cold phosphate-buffered saline. Tricolor analysis is performed with the tetramer-phycoerythrin, along with anti-CD8-Tricolor, and anti-CD38. The PBMCs are incubated with tetramer and antibodies on ice for 30 to 60 min and then washed twice before formaldehyde fixation. Gates are applied to contain >99.98% of control samples. Controls for the tetramers include both A*0201-negative individuals and A*0201-positive non-diseased donors. The percentage of cells stained with the tetramer is then determined by flow cytometry. The results indicate the number of cells in the PBMC sample that contain epitope-restricted CTLs, thereby readily indicating the extent of immune response to the 98P4B6 epitope, and thus the status of exposure to 98P4B6, or exposure to a vaccine that elicits a protective or therapeutic response.

Example 28 Use of Peptide Epitopes to Evaluate Recall Responses

The peptide epitopes of the invention are used as reagents to evaluate T cell responses, such as acute or recall responses, in patients. Such an analysis may be performed on patients who have recovered from 98P4B6-associated disease or who have been vaccinated with a 98P4B6 vaccine.

For example, the class I restricted CTL response of persons who have been vaccinated may be analyzed. The vaccine may be any 98P4B6 vaccine. PBMC are collected from vaccinated individuals and HLA typed. Appropriate peptide epitopes of the invention that, optimally, bear supermotifs to provide cross-reactivity with multiple HLA supertype family members, are then used for analysis of samples derived from individuals who bear that HLA type.

PBMC from vaccinated individuals are separated on Ficoll-Histopaque density gradients (Sigma Chemical Co., St. Louis, Mo.), washed three times in HBSS (GIBCO Laboratories), resuspended in RPMI-1640 (GIBCO Laboratories) supplemented with L-glutamine (2 mM), penicillin (50 U/ml), streptomycin (50 μg/ml), and Hepes (10 mM) containing 10% heat-inactivated human AB serum (complete RPMI) and plated using microculture formats. A synthetic peptide comprising an epitope of the invention is added at 10 μg/ml to each well and HBV core 128-140 epitope is added at 1 μg/ml to each well as a source of T cell help during the first week of stimulation.

In the microculture format, 4×10⁵ PBMC are stimulated with peptide in 8 replicate cultures in 96-well round bottom plate in 100 μl/well of complete RPMI. On days 3 and 10, 100 μl of complete RPMI and 20 U/ml final concentration of rIL-2 are added to each well. On day 7 the cultures are transferred into a 96-well flat-bottom plate and restimulated with peptide, rIL-2 and 10⁵ irradiated (3,000 rad) autologous feeder cells. The cultures are tested for cytotoxic activity on day 14. A positive CTL response requires two or more of the eight replicate cultures to display greater than 10% specific ⁵¹Cr release, based on comparison with non-diseased control subjects as previously described (Rehermann, et al., Nature Med. 2:1104, 1108, 1996; Rehermann et al., J. Clin. Invest. 97:1655-1665, 1996; and Rehermann et al. J. Clin. Invest. 98:1432-1440, 1996).

Target cell lines are autologous and allogeneic EBV-transformed B-LCL that are either purchased from the American Society for Histocompatibility and Immunogenetics (ASHI, Boston, Mass.) or established from the pool of patients as described (Guilhot, et al. J. Virol. 66:2670-2678, 1992).

Cytotoxicity assays are performed in the following manner. Target cells consist of either allogeneic HLA-matched or autologous EBV-transformed B lymphoblastoid cell line that are incubated overnight with the synthetic peptide epitope of the invention at 10 μM, and labeled with 100 Foci of ⁵¹Cr (Amersham Corp., Arlington Heights, Ill.) for 1 hour after which they are washed four times with HBSS.

Cytolytic activity is determined in a standard 4-h, split well ⁵¹Cr release assay using U-bottomed 96 well plates containing 3,000 targets/well. Stimulated PBMC are tested at effector/target (E/T) ratios of 20-50:1 on day 14. Percent cytotoxicity is determined from the formula: 100×[(experimental release−spontaneous release)/maximum release−spontaneous release)]. Maximum release is determined by lysis of targets by detergent (2% Triton X-100; Sigma Chemical Co., St. Louis, Mo.). Spontaneous release is <25% of maximum release for all experiments.

The results of such an analysis indicate the extent to which HLA-restricted CTL populations have been stimulated by previous exposure to 98P4B6 or a 98P4B6 vaccine.

Similarly, Class II restricted HTL responses may also be analyzed. Purified PBMC are cultured in a 96-well flat bottom plate at a density of 1.5×10⁵ cells/well and are stimulated with 10 μg/ml synthetic peptide of the invention, whole 98P4B6 antigen, or PHA. Cells are routinely plated in replicates of 4-6 wells for each condition. After seven days of culture, the medium is removed and replaced with fresh medium containing 10 U/ml IL-2. Two days later, 1 μCi ³H-thymidine is added to each well and incubation is continued for an additional 18 hours. Cellular DNA is then harvested on glass fiber mats and analyzed for ³H-thymidine incorporation. Antigen-specific T cell proliferation is calculated as the ratio of ³H-thymidine incorporation in the presence of antigen divided by the ³H-thymidine incorporation in the absence of antigen.

Example 29 Induction Of Specific CTL Response in Humans

A human clinical trial for an immunogenic composition comprising CTL and HTL epitopes of the invention is set up as an IND Phase I, dose escalation study and carried out as a randomized, double-blind, placebo-controlled trial. Such a trial is designed, for example, as follows:

A total of about 27 individuals are enrolled and divided into 3 groups:

Group I: 3 subjects are injected with placebo and 6 subjects are injected with 5 μg of peptide composition;

Group II: 3 subjects are injected with placebo and 6 subjects are injected with 50 μg peptide composition;

Group III: 3 subjects are injected with placebo and 6 subjects are injected with 500 μg of peptide composition.

After 4 weeks following the first injection, all subjects receive a booster inoculation at the same dosage.

The endpoints measured in this study relate to the safety and tolerability of the peptide composition as well as its immunogenicity. Cellular immune responses to the peptide composition are an index of the intrinsic activity of this the peptide composition, and can therefore be viewed as a measure of biological efficacy. The following summarize the clinical and laboratory data that relate to safety and efficacy endpoints.

Safety: The incidence of adverse events is monitored in the placebo and drug treatment group and assessed in terms of degree and reversibility.

Evaluation of Vaccine Efficacy: For evaluation of vaccine efficacy, subjects are bled before and after injection. Peripheral blood mononuclear cells are isolated from fresh heparinized blood by Ficoll-Hypaque density gradient centrifugation, aliquoted in freezing media and stored frozen. Samples are assayed for CTL and HTL activity.

The vaccine is found to be both safe and efficacious.

Example 30 Phase II Trials in Patients Expressing 98P4B6

Phase II trials are performed to study the effect of administering the CTL-HTL peptide compositions to patients having cancer that expresses 98P4B6. The main objectives of the trial are to determine an effective dose and regimen for inducing CTLs in cancer patients that express 98P4B6, to establish the safety of inducing a CTL and HTL response in these patients, and to see to what extent activation of CTLs improves the clinical picture of these patients, as manifested, e.g., by the reduction and/or shrinking of lesions. Such a study is designed, for example, as follows:

The studies are performed in multiple centers. The trial design is an open-label, uncontrolled, close escalation protocol wherein the peptide composition is administered as a single dose followed six weeks later by a single booster shot of the same dose. The dosages are 50, 500 and 5,000 micrograms per injection. Drug-associated adverse effects (severity and reversibility) are recorded.

There are three patient groupings. The first group is injected with 50 micrograms of the peptide composition and the second and third groups with 500 and 5,000 micrograms of peptide composition, respectively. The patients within each group range in age from 21-65 and represent diverse ethnic backgrounds. All of them have a tumor that expresses 98P4B6.

Clinical manifestations or antigen-specific T-cell responses are monitored to assess the effects of administering the peptide compositions. The vaccine composition is found to be both safe and efficacious in the treatment of 98P4B6 associated disease.

Example 31 Induction of CTL Responses Using a Prime Boost Protocol

A prime boost protocol similar in its underlying principle to that used to confirm the efficacy of a DNA vaccine in transgenic mice, such as described above in the Example entitled “The Plasmid Construct and the Degree to Which It Induces Immunogenicity,” can also be used for the administration of the vaccine to humans. Such a vaccine regimen can include an initial administration of, for example, naked DNA followed by a boost using recombinant virus encoding the vaccine, or recombinant protein/polypeptide or a peptide mixture administered in an adjuvant For example, the initial immunization may be performed using an expression vector, such as that constructed in the Example entitled “Construction of “Minigene” Multi-Epitope DNA Plasmids” in the form of naked nucleic acid administered IM (or SC or ID) in the amounts of 0.5-5 mg at multiple sites. The nucleic acid (0.1 to 1000 μg) can also be administered using a gene gun. Following an incubation period of 3-4 weeks, a booster dose is then administered. The booster can be recombinant fowlpox virus administered at a dose of 5-10⁷ to 5×10⁹ pfu. An alternative recombinant virus, such as an MVA, canarypox, adenovirus, or adeno-associated virus, can also be used for the booster, or the polyepitopic protein or a mixture of the peptides can be administered. For evaluation of vaccine efficacy, patient blood samples are obtained before immunization as well as at intervals following administration of the initial vaccine and booster doses of the vaccine. Peripheral blood mononuclear cells are isolated from fresh heparinized blood by Ficoll-Hypaque density gradient centrifugation, aliquoted in freezing media and stored frozen. Samples are assayed for CTL and HTL activity.

Analysis of the results indicates that a magnitude of response sufficient to achieve a therapeutic or protective immunity against 98P4B6 is generated.

Example 32 Administration of Vaccine Compositions Using Dendritic Cells (DC)

Vaccines comprising peptide epitopes of the invention can be administered using APCs, or “professional” APCs such as DC. In this example, peptide-pulsed DC are administered to a patient to stimulate a CTL response in vivo. In this method, dendritic cells are isolated, expanded, and pulsed with a vaccine comprising peptide CTL and HTL epitopes of the invention. The dendritic cells are infused back into the patient to elicit CTL and HTL responses in vivo. The induced CTL and HTL then destroy or facilitate destruction, respectively, of the target cells that bear the 98P4B6 protein from which the epitopes in the vaccine are derived.

For example, a cocktail of epitope-comprising peptides is administered ex vivo to PBMC, or isolated DC therefrom. A pharmaceutical to facilitate harvesting of DC can be used, such as Progenipoietin™ (Monsanto, St. Louis, Mo.) or GM-CSF/IL-4. After pulsing the DC with peptides, and prior to reinfusion into patients, the DC are washed to remove unbound peptides.

As appreciated clinically, and readily determined by one of skill based on clinical outcomes, the number of DC reinfused into the patient can vary (see, e.g., Nature Med. 4:328, 1998; Nature Med. 2:52, 1996 and Prostate 32:272, 1997). Although 2-50×10⁶ DC per patient are typically administered, larger number of DC, such as 10⁷ or 10⁸ can also be provided. Such cell populations typically contain between 50-90% DC.

In some embodiments, peptide-loaded PBMC are injected into patients without purification of the DC. For example, PBMC generated after treatment with an agent such as Progenipoietin™ are injected into patients without purification of the DC. The total number of PBMC that are administered often ranges from 10⁸ to 101°. Generally, the cell doses injected into patients is based on the percentage of DC in the blood of each patient, as determined, for example, by immunofluorescence analysis with specific anti-DC antibodies. Thus, for example, if Progenipoietin™ mobilizes 2% DC in the peripheral blood of a given patient, and that patient is to receive 5×10⁶ DC, then the patient will be injected with a total of 2.5×10⁸ peptide-loaded PBMC. The percent DC mobilized by an agent such as Progenipoietin™ is typically estimated to be between 2-10%, but can vary as appreciated by one of skill in the art.

Ex Vivo Activation of CTL/HTL Responses

Alternatively, ex vivo CTL or HTL responses to 98P4B6 antigens can be induced by incubating, in tissue culture, the patient's, or genetically compatible, CTL or HTL precursor cells together with a source of APC, such as DC, and immunogenic peptides. After an appropriate incubation time (typically about 7-28 days), in which the precursor cells are activated and expanded into effector cells, the cells are infused into the patient, where they will destroy (CTL) or facilitate destruction (HTL) of their specific target cells, i.e., tumor cells.

Example 33 An Alternative Method of Identifying and Confirming Motif-Bearing Peptides

Another method of identifying and confirming motif-bearing peptides is to elute them from cells bearing defined MHC molecules. For example, EBV transformed B cell lines used for tissue typing have been extensively characterized to determine which HLA molecules they express. In certain cases these cells express only a single type of HLA molecule. These cells can be transfected with nucleic acids that express the antigen of interest, e.g. 98P4B6. Peptides produced by endogenous antigen processing of peptides produced as a result of transfection will then bind to HLA molecules within the cell and be transported and displayed on the cell's surface. Peptides are then eluted from the HLA molecules by exposure to mild acid conditions and their amino acid sequence determined, e.g., by mass spectral analysis (e.g., Kubo et al., J. Immunol. 152:3913, 1994). Because the majority of peptides that bind a particular HLA molecule are motif-bearing, this is an alternative modality for obtaining the motif-bearing peptides correlated with the particular HLA molecule expressed on the cell.

Alternatively, cell lines that do not express endogenous HLA molecules can be transfected with an expression construct encoding a single HLA allele. These cells can then be used as described, i.e., they can then be transfected with nucleic acids that encode 98P4B6 to isolate peptides corresponding to 98P4B6 that have been presented on the cell surface. Peptides obtained from such an analysis will bear motif(s) that correspond to binding to the single HLA allele that is expressed in the cell.

As appreciated by one in the art, one can perform a similar analysis on a cell bearing more than one HLA allele and subsequently determine peptides specific for each HLA allele expressed. Moreover, one of skill would also recognize that means other than transfection, such as loading with a protein antigen, can be used to provide a source of antigen to the cell.

Example 34 Complementary Polynucleotides

Sequences complementary to the 98P4B6-encoding sequences, or any parts thereof, are used to detect, decrease, or inhibit expression of naturally occurring 98P4B6. Although use of oligonucleotides comprising from about 15 to 30 base pairs is described, essentially the same procedure is used with smaller or with larger sequence fragments. Appropriate oligonucleotides are designed using, e.g., OLIGO 4.06 software (National Biosciences) and the coding sequence of 98P4B6. To inhibit transcription, a complementary oligonucleotide is designed from the most unique 5′ sequence and used to prevent promoter binding to the coding sequence. To inhibit translation, a complementary oligonucleotide is designed to prevent ribosomal binding to a 98P4B6-encoding transcript.

Example 35 Purification of Naturally-Occurring or Recombinant 98P4B6 Using 98P4B6-Specific Antibodies

Naturally occurring or recombinant 98P4B6 is substantially purified by immunoaffinity chromatography using antibodies specific for 98P4B6. An immunoaffinity column is constructed by covalently coupling anti-98P4B6 antibody to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Pharmacia Biotech). After the coupling, the resin is blocked and washed according to the manufacturers instructions.

Media containing 98P4B6 are passed over the immunoaffinity column, and the column is washed under conditions that allow the preferential absorbance of 98P4B6 (e.g., high ionic strength buffers in the presence of detergent). The column is eluted under conditions that disrupt antibody/98P4B6 binding (e.g., a buffer of pH 2 to pH 3, or a high concentration of a chaotrope, such as urea or thiocyanate ion), and GCR.P is collected.

Example 36 Identification of Molecules which Interact with 98P4B6

98P4B6, or biologically active fragments thereof, are labeled with 121 1 Bolton-Hunter reagent. (See, e.g., Bolton et al. (1973) Biochem. J. 133:529.) Candidate molecules previously arrayed in the wells of a multi-well plate are incubated with the labeled 98P4B6, washed, and any wells with labeled 98P4B6 complex are assayed. Data obtained using different concentrations of 98P4B6 are used to calculate values for the number, affinity, and association of 98P4B6 with the candidate molecules.

Example 37 In Vivo Assay for 98P4B6 Tumor Growth Promotion

The effect of the 98P4B6 protein on tumor cell growth is evaluated in vivo by gene overexpression in tumor-bearing mice. For example, prostate (PC3), lung (A427), stomach, ovarian (PA1) and uterus cell lines are engineered to express 98P4B6. SCID mice are injected subcutaneously on each flank with 1×10⁶ of PC3, A427, PA1, or NIH-3T3 cells containing tkNeo empty vector or 98P4B6. At least two strategies may be used: (1) Constitutive 98P4B6 expression under regulation of a promoter such as a constitutive promoter obtained from the genomes of viruses such as polyoma virus, fowlpox virus (UK 2,211,504 published 5 Jul. 1989), adenovirus (such as Adenovirus 2), bovine papiloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus, and Simian Virus 40 (SV40), or from heterologous mammalian promoters, e.g., the actin promoter or an immunoglobulin promoter, provided such promoters are compatible with the host cell systems, and (2) Regulated expression under control of an inducible vector system, such as ecdysone, tet, etc., provided such promoters are compatible with the host cell systems. Tumor volume is then monitored at the appearance of palpable tumors and followed over time to determine if 98P4B6-expressing cells grow at a faster rate and whether tumors produced by 98P4B6-expressing cells demonstrate characteristics of altered aggressiveness (e.g. enhanced metastasis, vascularization, reduced responsiveness to chemotherapeutic drugs).

Additionally, mice can be implanted with 1×10⁵ of the same cells, orthotopically to determine if 98P4B6 has an effect on local growth in the prostate or on the ability of the cells to metastasize, specifically to lungs, lymph nodes, and bone marrow.

The assay is also useful to determine the 98P4B6 inhibitory effect of candidate therapeutic compositions, such as for example, 98P4B6 intrabodies, 98P4B6 antisense molecules and ribozymes.

Example 38 98P4B6 Monoclonal Antibody-mediated Inhibition of Tumors In Vivo

The significant expression of 98P4B6 in prostate, lung, stomach, ovary, and uterus cancer tissues, its restrictive expression in normal tissues, together with its expected cell surface expression makes 98P4B6 an excellent target for antibody therapy. Similarly, 98P4B6 is a target for T-cell based immunotherapy. Thus, the therapeutic efficacy of anti-P4B6 mAbs in human prostate cancer xenograft mouse models is evaluated by using androgen-independent LAPC-4 and LAPC-9 xenografts (Craft, N., et al., Cancer Res, 1999. 59(19): p. 5030-6) and the androgen independent recombinant cell line PC3-98P4B6 (see, e.g., Kaighn, M. E., et al., Invest Urol, 1979. 17(1): p. 16-23). Similar approaches using patient derived xenografts or xenograft cell lines are used for cancers listed in Table I.

Antibody efficacy on tumor growth and metastasis formation is studied, e.g., in a mouse orthotopic prostate cancer xenograft models and mouse lung, uterus, or stomach xenograft models. The antibodies can be unconjugated, as discussed in this Example, or can be conjugated to a therapeutic modality, as appreciated in the art. Anti-98P4B6 mAbs inhibit formation of both the androgen-dependent LAPC-9 and androgen-independent PC3-98P4B6 tumor xenografts. Anti-98P4B6 mAbs also retard the growth of established orthotopic tumors and prolonged survival of tumor-bearing mice. These results indicate the utility of anti-98P4B6 mAbs in the treatment of local and advanced stages of cancer. (See, e.g., (Saffran, D., et al., PNAS 10:1073-1078).

Administration of the anti-98P4B6 mAbs can lead to retardation of established orthotopic tumor growth and inhibition of metastasis to distant sites, resulting in a significant prolongation in the survival of tumor-bearing mice. These studies indicate that 98P4B6 is an attractive target for immunotherapy and demonstrate the therapeutic potential of anti-98P4B6 mAbs for the treatment of local and metastatic cancer. This example demonstrates that unconjugated 98P4B6 monoclonal antibodies are effective to inhibit the growth of human prostate tumor xenografts, as well as lung, uterus, or stomach xenograft grown in SCID mice; accordingly a combination of such efficacious monoclonal antibodies is also effective.

Tumor inhibition using multiple unconjugated 98P4B6 mAbs

Materials and Methods

98P4B6 Monoclonal Antibodies:

Monoclonal antibodies are raised against 98P4B6 as described in Example 11 entitled “Generation of 98P4B6 Monoclonal Antibodies (mAbs).” The antibodies are characterized by ELISA, Western blot, FACS, and immunoprecipitation for their capacity to bind 98P4B6. Epitope mapping data for the anti-98P4B6 mAbs, as determined by ELISA and Western analysis, recognize epitopes on the 98P4B6 protein. Immunohistochemical analysis of cancer tissues and cells with these antibodies is performed.

The monoclonal antibodies are purified from ascites or hybridoma tissue culture supernatants by Protein-G Sepharose chromatography, dialyzed against PBS, filter sterilized, and stored at −20° C. Protein determinations are performed by a Bradford assay (Bio-Rad, Hercules, Calif.). A therapeutic monoclonal antibody or a cocktail comprising a mixture of individual monoclonal antibodies is prepared and used for the treatment of mice receiving subcutaneous or orthotopic injections of LAPC-9 tumor xenografts.

Cancer Xenografts and Cell Lines

The LAPC-9 xenograft, which expresses a wild-type androgen receptor and produces prostate-specific antigen (PSA), is passaged in 6- to 8-week-old male ICR-severe combined immunodeficient (SCID) mice (Taconic Farms) by s.c. trocar implant (Craft, N., et al., supra). The prostate (PC3), lung (A427), ovarian (PA1) carcinoma cell lines (American Type Culture Collection) are maintained in RPMI or DMEM supplemented with L-glutamine and 10% FBS.

PC3-98P4B6, A427-98P4B6, PA1-98P4B6 and 3T3-98P4B6 cell populations are generated by retroviral gene transfer as described in Hubert, R. S., et al., STEAP: a prostate-specific cell-surface antigen highly expressed in human prostate tumors. Proc Natl Acad Sd USA, 1999. 96(25): p. 14523-8. Anti-98P4B6 staining is detected by using an FITC-conjugated goat anti-mouse antibody (Southern Biotechnology Associates) followed by analysis on a Coulter Epics-XL flow cytometer.

Xenograft Mouse Models.

Subcutaneous (s.c.) tumors are generated by injection of 1×10⁶ LAPC-9, PC3, PC3-98P4B6, A427, A427-98P4B6, PA1, PA1-98P4B6, 3T3 or 3T3-98P4B6 cells mixed at a 1:1 dilution with Matrigel (Collaborative Research) in the right flank of male SCID mice. To test antibody efficacy on tumor formation, i.p. antibody injections are started on the same day as tumor-cell injections. As a control, mice are injected with either purified mouse IgG (ICN) or PBS; or a purified monoclonal antibody that recognizes an irrelevant antigen not expressed in human cells. In preliminary studies, no difference is found between mouse IgG or PBS on tumor growth. Tumor sizes are determined by vernier caliper measurements, and the tumor volume is calculated as length×width×height. Mice with s.c. tumors greater than 1.5 cm in diameter are sacrificed. PSA levels are determined by using a PSA ELISA kit (Anogen, Mississauga, Ontario). Circulating levels of anti-98P4B6 mAbs are determined by a capture ELISA kit (Bethyl Laboratories, Montgomery, Tex.). (See, e.g., (Saffran, D., et al., PNAS 10:1073-1078).

Orthotopic injections are performed under anesthesia by using ketamine/xylazine. For prostate orthotopic studies, an incision is made through the abdominal muscles to expose the bladder and seminal vesicles, which then are delivered through the incision to expose the dorsal prostate. LAPC-9 or PC3 cells (5×10⁵) mixed with Mabigel are injected into each dorsal lobe in a 10-μl volume. To monitor tumor growth, mice are bled on a weekly basis for determination of PSA levels. The mice are segregated into groups for the appropriate treatments, with anti-98P4B6 or control mAbs being injected i.p.

Anti-98P4B6 mAbs Inhibit Growth of 98P4B6-Expressing Xenograft-Cancer Tumors

The effect of anti-98P4B6 mAbs on tumor formation is tested by using LAPC-9 and PC3-98P4B6 orthotopic models. As compared with the s.c. tumor model, the orthotopic model, which requires injection of tumor cells directly in the mouse prostate, lung, or ovary, respectively, results in a local tumor growth, development of metastasis in distal sites, deterioration of mouse health, and subsequent death (Saffran, D., et al., PNAS supra; Fu, X., et al., Int J Cancer, 1992. 52(6): p. 987-90; Kubota, T., J Cell Biochem, 1994. 56(1): p. 4-8). The features make the orthotopic model more representative of human disease progression and allowed us to follow the therapeutic effect of mAbs on clinically relevant end points.

Accordingly, tumor cells are injected into the mouse prostate, lung, or ovary, and 2 days later, the mice are segregated into two groups and treated with either: a) 200-500 μg, of anti-98P4B6 Ab, or b) PBS three times per week for two to five weeks.

A major advantage of the orthotopic cancer model is the ability to study the development of metastases. Formation of metastasis in mice bearing established orthotopic tumors is studies by IHC analysis on lung sections using an antibody against a prostate-specific cell-surface protein STEAP expressed at high levels in LAPC-9 xenografts (Hubert, R. S., et al., Proc Natl Acad Sci USA, 1999. 96(25): p. 14523-8).

Mice bearing established orthotopic LAPC-9 or PC3-98P4B6 tumors are administered 1000 μg injections of either anti-98P4B6 mAb or PBS over a 4-week period. Mice in both groups are allowed to establish a high tumor burden (PSA levels greater than 300 ng/ml for IAPC-9), to ensure a high frequency of metastasis formation in mouse lungs. Mice then are killed and their prostate and lungs are analyzed for the presence of tumor cells by IHC analysis.

These studies demonstrate a broad anti-tumor efficacy of anti-98P4B6 antibodies on initiation and progression of prostate cancer in xenograft mouse models. Anti-98P4B6 antibodies inhibit tumor formation of both androgen-dependent and androgen-independent tumors as well as retarding the growth of already established tumors and prolong the survival of treated mice. Moreover, anti-98P4B6 mAbs demonstrate a dramatic inhibitory effect on the spread of local prostate tumor to distal sites, even in the presence of a large tumor burden. Thus, anti-98P4B6 mAbs are efficacious on major clinically relevant end points (tumor growth), prolongation of survival, and health.

Example 39 Therapeutic and Diagnostic use of Anti-98P4B6 Antibodies in Humans

Anti-98P4B6 monoclonal antibodies are safely and effectively used for diagnostic, prophylactic, prognostic and/or therapeutic purposes in humans. Western blot and immunohistochemical analysis of cancer tissues and cancer xenografts with anti-98P4B6 mAb show strong extensive staining in carcinoma but significantly lower or undetectable levels in normal tissues. Detection of 98P4B6 in carcinoma and in metastatic disease demonstrates the usefulness of the mAb as a diagnostic and/or prognostic indicator. Anti-98P4B6 antibodies are therefore used in diagnostic applications such as immunohistochemistry of kidney biopsy specimens to detect cancer from suspect patients.

As determined by flow cytometry, anti-98P4B6 mAb specifically binds to carcinoma cells. Thus, anti-98P4B6 antibodies are used in diagnostic whole body imaging applications, such as radioimmunoscintgraphy and radioimmunotherapy, (see, e.g., Potamianos S., et. al. Anticancer Res 20(2A):925-948 (2000)) for the detection of localized and metastatic cancers that exhibit expression of 98P4B6. Shedding or release of an extracellular domain of 98P4B6 into the extracellular milieu, such as that seen for alkaline phosphodiesterase B10 (Meerson, N. R., Hepatology 27:563-568 (1998)), allows diagnostic detection of 98P4B6 by anti-98P4B6 antibodies in serum and/or urine samples from suspect patients.

Anti-98P4B6 antibodies that specifically bind 98P4B6 are used in therapeutic applications for the treatment of cancers that express 98P4B6. Anti-98P4B6 antibodies are used as an unconjugated modality and as conjugated form in which the antibodies are attached to one of various therapeutic or imaging modalities well known in the art, such as a prodrugs, enzymes or radioisotopes. In preclinical studies, unconjugated and conjugated anti-98P4B6 antibodies are tested for efficacy of tumor prevention and growth inhibition in the SCID mouse cancer xenograft models, e.g., kidney cancer models AGS-K3 and AGS-K6, (see, e.g., the Example entitled “98P4B6 Monoclonal Antibody-mediated Inhibition of Bladder and Lung Tumors In Vivo”). Either conjugated and unconjugated anti-98P4B6 antibodies are used as a therapeutic modality in human clinical trials either alone or in combination with other treatments as described in following Examples.

Example 40 Human Clinical Trials for the Treatment and Diagnosis of Human Carcinomas through use of Human Anti-98P4B6 Antibodies In Vivo

Antibodies are used in accordance with the present invention which recognize an epitope on 98P4B6, and are used in the treatment of certain tumors such as those listed in Table I. Based upon a number of factors, including 98P4B6 expression levels, tumors such as those listed in Table I are presently preferred indications. In connection with each of these indications, three clinical approaches are successfully pursued.

I.) Adjunctive therapy: In adjunctive therapy, patients are treated with anti-98P4B6 antibodies in combination with a chemotherapeutic or antineoplastic agent and/or radiation therapy. Primary cancer targets, such as those listed in Table I, are treated under standard protocols by the addition anti-98P4B6 antibodies to standard first and second line therapy. Protocol designs address effectiveness as assessed by reduction in tumor mass as well as the ability to reduce usual doses of standard chemotherapy. These dosage reductions allow additional and/or prolonged therapy by reducing dose-related toxicity of the chemotherapeutic agent. Anti-98P4B6 antibodies are utilized in several adjunctive clinical trials in combination with the chemotherapeutic or antineoplastic agents adriamycin (advanced prostrate carcinoma), cisplatin (advanced head and neck and lung carcinomas), taxol (breast cancer), and doxorubicin (preclinical).

II.) Monotherapy: In connection with the use of the anti-98P4B6 antibodies in monotherapy of tumors, the antibodies are administered to patients without a chemotherapeutic or antineoplastic agent. In one embodiment, monotherapy is conducted clinically in end stage cancer patients with extensive metastatic disease. Patients show some disease stabilization. Trials demonstrate an effect in refractory patients with cancerous tumors.

III.) Imaging Agent: Through binding a radionuclide (e.g., iodine or yttrium (I¹³¹, Y⁹⁰) to anti-98P4B6 antibodies, the radiolabeled antibodies are utilized as a diagnostic and/or imaging agent. In such a role, the labeled antibodies localize to both solid tumors, as well as, metastatic lesions of cells expressing 98P4B6. In connection with the use of the anti-98P4B6 antibodies as imaging agents, the antibodies are used as an adjunct to surgical treatment of solid tumors, as both a pre-surgical screen as well as a post-operative follow-up to determine what tumor remains and/or returns. In one embodiment, a (¹¹¹In)-98P4B6 antibody is used as an imaging agent in a Phase I human clinical trial in patients having a carcinoma that expresses 98P4B6 (by analogy see, e.g., Divgi et al. J. Natl. Cancer Inst. 83:97-104 (1991)). Patients are followed with standard anterior and posterior gamma camera. The results indicate that primary lesions and metastatic lesions are identified

Dose and Route of Administration

As appreciated by those of ordinary skill in the art, dosing considerations can be determined through comparison with the analogous products that are in the clinic. Thus, anti-98P4B6 antibodies can be administered with doses in the range of 5 to 400 mg/m², with the lower doses used, e.g., in connection with safety studies. The affinity of anti-98P4B6 antibodies relative to the affinity of a known antibody for its target is one parameter used by those of skill in the art for determining analogous dose regimens. Further, ant-98P4B6 antibodies that are fully human antibodies, as compared to the chimeric antibody, have slower clearance; accordingly, dosing in patients with such fully human anti-98P4B6 antibodies can be lower, perhaps in the range of 50 to 300 mg/m², and still remain efficacious. Dosing in mg/m², as opposed to the conventional measurement of dose in mg/kg, is a measurement based on surface area and is a convenient dosing measurement that is designed to include patients of all sizes from infants to adults.

Three distinct delivery approaches are useful for delivery of anti-98P4B6 antibodies. Conventional intravenous delivery is one standard delivery technique for many tumors. However, in connection with tumors in the peritoneal cavity, such as tumors of the ovaries, biliary duct, other ducts, and the like, intraperitoneal administration may prove favorable for obtaining high dose of antibody at the tumor and to also minimize antibody clearance. In a similar manner, certain solid tumors possess vasculature that is appropriate for regional perfusion. Regional perfusion allows for a high dose of antibody at the site of a tumor and minimizes short term clearance of the antibody.

Clinical Development Plan (CDP)

Overview: The CDP follows and develops treatments of anti-98P4B6 antibodies in connection with adjunctive therapy, monotherapy, and as an imaging agent. Trials initially demonstrate safety and thereafter confirm efficacy in repeat doses. Trails are open label comparing standard chemotherapy with standard therapy plus anU-98P4B6 antibodies. As will be appreciated, one criteria that can be utilized in connection with enrollment of patients is 98P4B6 expression levels in their tumors as determined by biopsy.

As with any protein or antibody infusion-based therapeutic, safety concerns are related primarily to (i) cytokine release syndrome, i.e., hypotension, fever, shaking, chills; (ii) the development of an immunogenic response to the material (i.e., development of human antibodies by the patient to the antibody therapeutic, or HAHA response); and, (iii) toxicity to normal cells that express 98P4B6. Standard tests and follow-up are utilized to monitor each of these safety concerns. Anti-98P4B6 antibodies are found to be safe upon human administration.

Example 41 Human Clinical Trial Adjunctive Therapy with Human Anti-98P4B6 Antibody and Chemotherapeutic Agent

A phase I human clinical trial is initiated to assess the safety of six intravenous doses of a human anti-98P4B6 antibody in connection with the treatment of a solid tumor, e.g., a cancer of a tissue listed in Table I. In the study, the safety of single doses of anti-98P486 antibodies when utilized as an adjunctive therapy to an antineoplastic or chemotherapeutic agent as defined herein, such as, without limitation: cisplatin, topotecan, doxorubicin, adriamycin, taxol, or the like, is assessed. The trial design includes delivery of six single doses of an anti-98P4B6 antibody with dosage of antibody escalating from approximately about 25 mg/m² to about 275 mg/m² over the course of the treatment in accordance with the following schedule:

Day 0 Day 7 Day 14 Day 21 Day 28 Day 35 mAb Dose 25 mg/m² 75 mg/m² 125 mg/m² 175 mg/m² 225 mg/m² 275 mg/m² Chemotherapy + + + + + + (standard dose)

Patients are closely followed for one-week following each administration of antibody and chemotherapy. In particular, patients are assessed for the safety concerns mentioned above: (i) cytokine release syndrome, i.e., hypotension, fever, shaking, chills; (ii) the development of an immunogenic response to the material (i.e., development of human antibodies by the patient to the human antibody therapeutic, or HAHA response); and, (iii) toxicity to normal cells that express 98P4B6. Standard tests and follow-up are utilized to monitor each of these safety concerns. Patients are also assessed for clinical outcome, and particularly reduction in tumor mass as evidenced by MRI or other imaging.

The anti-98P4B6 antibodies are demonstrated to be safe and efficacious, Phase II trials confirm the efficacy and refine optimum dosing.

Example 42 Human Clinical Trial: Monotherapy with Human Anti-98P4B6 Antibody

Anti-98P4B6 antibodies are safe in connection with the above-discussed adjunctive trial, a Phase II human clinical trial confirms the efficacy and optimum dosing for monotherapy. Such trial is accomplished, and entails the same safety and outcome analyses, to the above-described adjunctive trial with the exception being that patients do not receive chemotherapy concurrently with the receipt of doses of anti-98P4B6 antibodies.

Example 43 Human Clinical Trial: Diagnostic Imaging with Anti-98P4B6 Antibody

Once again, as the adjunctive therapy discussed above is safe within the safety criteria discussed above, a human clinical trial is conducted concerning the use of anti-98P4B6 antibodies as a diagnostic imaging agent. The protocol is designed in a substantially similar manner to those described in the art, such as in Divgi et al. J. Natl. Cancer Inst. 83:97-104 (1991). The antibodies are found to be both safe and efficacious when used as a diagnostic modality.

Example 44 Homology Comparison of 98P4B6 to Known Sequences

The 98P4B6 gene is homologous to a cloned and sequenced gene, namely human STAMP1 (gi 15418732) (Korkmaz, K. S et al., J. Biol. Chem. 2002, 277: 36689), showing 99% identity and 99% homology to that gene (FIG. 4). The 98P4B6 protein also shows 99% identity and 99% homology to another human six transmembrane epithelial antigen of prostate 2 (gi 23308593) (Walker, M. G et al., Genome Res. 1999, 9: 1198; Porkka, K. P., Helenius, M. A. and Visakorpi, T, Lab. Invest. 2002, 82: 1573). The closest mouse homolog to 98P4B6 is six transmembrane epithelial antigen of prostate 2 (gi 28501136), with 97% identity and 99% homology. We have identified several variants of the 98P4B6 protein, including 4 splice variants and 3 SNPs (FIG. 11). The 98P4B6 v.1 protein consists of 454 amino acids, with calculated molecular weight of 52 kDa, and pl of 8.7. It is a 6 transmembrane protein that can localize to the cell surface or possibly to the endoplasmic reticulum (Table VI). Several 98P4B6 variants, including v.1, v.5-8, v.13, v.14, v.21, v.25 share similar features, such protein motifs with functional significance, as well as structural commonalties such as multiple transmembrane domains. The 98P4B6 v.2 is a short protein with no known motifs.

Motif analysis revealed the presence of several known motifs, including oxido-reductase, homocysteine hydrolase and dudulin motifs. Variant v.7 and SNPs of this variant also carry an Ets motif, often associated with transcriptional activity.

Several oxidoreductases have been identified in mammalian cells, including the NADH/quinone oxidoreductase. This protein associate with the cell membrane and function as a proton/Na+ pump, which regulates the protein degradation of the tumor suppressor p53, and protects mammalian cells from oxidative stress, cytotoxicity, and mutages (Asher G, et al., Proc Natl Acad Sci USA. 2002, 99:13125; Jaiswal A K, Arch Biochem Biophys 2000, 375:62 Yano T, Mol Aspects Med 2002, 23:345). Homocysteine hydrolase is an enzyme known to catalyze the breakdown of S-adenosylhomocysteine to homocysteine and adenosine, ultimately regulating trans-methylation, thereby regulating protein expression, cell cycle and proliferation (Turner M A et al; Cell Biochem Biophys 2000; 33:101; Zhang et al, J Biol. Chem. 2001; 276:35867)

This information indicates that 98P4B6 plays a role in the cell growth of mammalian cells, regulate gene transcription and transport of electrons and small molecules. Accordingly, when 98P4B6 functions as a regulator of cell growth, tumor formation, or as a modulator of transcription involved in activating genes associated with inflammation, tumorigenesis, or proliferation, 98P4B6 is used for therapeutic, diagnostic, prognostic and/or preventative purposes. In addition, when a molecule, such as a variant or polymorphism of 98P4B6 is expressed in cancerous tissues, it is used for therapeutic, diagnostic, prognostic and/or preventative purposes.

Example 45 Phenotypic Effects of STEAP-2 Expression

Experiments regarding the expression of STEAP-2 protein having the amino acid sequence shown in FIG. 2 and encoded by a cDNA insert in a plasmid deposited with the American Type Culture Collection on 2 Jul. 1999 and assigned as ATCC Accession No. PTA-311. As deduced from the coding sequence, the open reading frame encodes 454 amino acids with 6 transmembrane domains. A summary of the characteristics associated with STEAP-2 protein is shown on FIG. 19.

This deposit was made under the provisions of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purpose of Patient Procedure and the Regulations thereunder (Budapest Treaty). This assures maintenance of a viable culture of the deposit for 30 years from the date of deposit and for at least five (5) years after the most recent request for the furnishing of a sample of the deposit received by the depository. The deposits will be made available by ATCC under the terms of the Budapest Treaty, and subject to an agreement between Genentech, Inc. and ATCC, which assures that all restrictions imposed by the depositor on the availability to the public of the deposited material will be irrevocably removed upon the granting of the pertinent U.S. patent, assures permanent and unrestricted availability of the progeny of the culture of the deposit to the public upon issuance of the pertinent U.S. patent or upon laying open to the public of any U.S. or foreign patent application, whichever comes first, and assures availability of the progeny to one determined by the U.S. Commissioner of Patents and Trademarks to be entitled thereto according to 35 USC §122 and the Commissioners rules pursuant thereto (including 37 CFR §1.14 with particular reference to 886 OG 638).

The assignee of the present application has agreed that if a culture of the materials on deposit should die or be lost or destroyed when cultivated under suitable conditions, the materials will be promptly replaced on notification with another of the same. Availability of the deposited material is not to be construed as a license to practice the invention in contravention of the rights granted under the authority of any government in accordance with its patent laws.

The data set forth in the present patent application provide an expression profile of the STEAP-2 protein that is predominantly specific for the prostate among normal tissues, for certain types of prostate tumors as well as other tumors. This evidence is based on detecting messenger RNA using Northern blotting. In keeping with standard practice in this industry, Northern blots are routinely used to assess gene expression, as it does not require the time consuming process of synthesizing the relevant protein, raising antibodies, assuring the specificity of the antibodies, required for Western blotting of proteins and the histological examination of tissues. Northern blotting offers a credible and efficient method of assessing RNA expression and expression levels.

This Example demonstrates that STEAP-2 protein is, indeed, produced. In summary, the experiments show that PC-3 cells and 3T3 cells which were modified to contain an expression system for STEAP-2 showed enhanced levels of tyrosine phosphorylation in general, and of phosphorylation of ERK protein in particular. The data also show that PC-3 cells that contain an expression system for STEAP-2 showed modified calcium flux, a modified response to paclitaxel, and a general inhibition of drug-induced apoptosis. These are effects exhibited at the protein level, thus these data alone are probative that the STEAP-2 protein exists.

Furthermore, although such phenotypic effects are protein-mediated, further evidence indicates that the STEAP-2 protein itself is the mediator of the effects. This evidence is obtained by utilizing a modified STEAP-2 protein. An expression system is stably introduced into PC3 and 3T3 cells which allows the expression of a modified form of STEAP-2, designated STEAP-2CFl, where “Fl” stands for flag. STEAP-2CFI is a STEAP-2 protein having a peptide extension, i.e., a Flag epitope that alters the physical conformation of this protein. The Flag epitope is a string 8 amino acids, often introduced at either the amino or carboxy termini of protein as a means of identifying and following a recombinant protein in engineered cells (Slootstra J W et al., Mol Divers 1997, 2:156). In most cases, the introduction of the Flag epitope at either termini of a protein has little effect on the natural function and location of that protein (Molloy S S et al., EMBO J. 1994, 13:18). However, this is dependent on the characteristics of the protein being Flag tagged. Recent studies have shown that a Flag tag affects the function and conformation of select proteins such as the CLN3 protein (see, e.g., Haskell R E, et al. Mol Genet Metab 1999, 66:253). As with CLN3, introducing a Flag epitope tag to the C-terminus of STEAP-2 alters the physical conformation and properties of this protein. Altering the STEAP-2 protein with the C-Flag epitope resulted in a significant decrease in the effects otherwise observed, including phosphorylation of ERK and resistance to drug-induced cell death. The data indicate that it is the STEAP-2 protein that mediated these phenotypic effects. Finally, in vitro translation studies using rabbit reticulocyte lysate, showed that the STEAP-2 protein is translated and exhibits the expected molecular weight.

FIGS. 20 and 21 show the results obtained when PC-3 and 3T3 cells, respectively, were modified to contain the retroviral expression system pSR□ encoding the indicated proteins, including STEAP-1, STEAP-2 and STEAP-2CFI, respectively. Gene-specific protein expression was driven from a long terminal repeat (LTR), and the Neomycin resistance gene was used for selection of mammalian cells that stably express the protein. PC-3 ahd 3T3 cells were transduced with the retrovirus, selected in the presence of G418 and cultured under conditions which permit expression of the STEAP-2 coding sequence. The cells were grown overnight in low concentrations of FBS (0.5-1% FBS) and were then stimulated with 10% FBS. The cells were lysed in RIPA buffer and quantitated for protein concentration. Whole cell lysates were separated by SDS-PAGE and analyzed by Western blotting using anti-phospho-ERK (Cell Signaling Inc.) or anti-phosphotyrosine (UBI) antibodies (FIGS. 20, 21, and 22). As shown on FIG. 20, as compared to untransformed PC-3 cells, cells modified to contain STEAP-2 contain enhanced amounts of phosphorylated tyrosine. Similar results from an analogous experiment on 3T3 cells are shown on page 3. In this latter experiment, the STEAP-2CFI expression system was also transfected into 3T3 cells, which cells were used as a control. As shown on FIG. 21, the enhanced-phosphorylation found in the presence of native STEAP-2 was significantly reduced when the conformation of the protein was altered. These results thus show conclusively that the STEAP-2 protein was produced and mediated the above-described phenotypic effects.

FIG. 22 shows similar results, both in PC-3 and 3T3 cells where phosphorylation of ERK, specifically, is detected. The protocol is similar to that set forth in paragraph 5 above, except that rather than probing the gels with antibodies specific for phosphotyrosine the gels were probed both the anti-ERK and anti-phospho-ERK antibodies. As shown on FIG. 22, in the presence of 10% FBS, both PC-3 cells and 3T3 cells modified to express STEAP-2 showed phosphorylation of ERK which was not detectable in cells transformed to contain STEAP-2CFI. In contrast to control PC-3 cells which exhibit no background ERK phosphorylation, control 3T3-neo cells show low levels of endogenous ERK phosphorylation. Treatment with 10% FBS enhanced phosphorylation of ERK protein in cells expressing STEAP-2 relative to 3T3-neo cells, while no increase in ERK phosphorylation was observed in 3T3 cells expressing modified STEAP-2, i.e. STEAP-2 CFI.

Other effects on cellular metabolism in cells modified to contain a STEAP-2 expression system were also shown in our data. FIG. 23 shows that when cells with and without expression systems for STEAP-2 were measured for calcium flux in the presence of LPA, calcium flux was enhanced in the STEAP-2 containing cells. Using FACS analysis and commercially available indicators (Molecular Probes), parental cells and cells expressing STEAP-2 were compared for their ability to transport calcium. PC3-neo and PC3-STEAP-2 cells were loaded with calcium responsive indicators Fluo4 and Fura red, incubated in the presence or absence of calcium and LPA, and analyzed by flow cytometry. PC3 cells expressing a known calcium transporter, PC3-83P3H3 pCaT were used as positive control (Biochem Biophys Res Commun. 2001, 282:729). The table on FIG. 23 shows that STEAP72 mediates calcium flux in response to LPA, and that the magnitude of calcium flux is comparable to that produced by a known calcium channel.

In addition, STEAP-2 expressing PC3 cells demonstrated increased sensitivity to agatoxin, a calcium channel blocker as compared to PC3-neo cells. These results indicate that STEAP-2 expression renders PC3 cells sensitive to treatment with the Ca++ channel inhibitors. Information derived from the above experiments provides a mechanism by which cancer cells are regulated. This is particularly relevant in the case of calcium, as calcium channel inhibitors have been reported to induce the death of certain cancer cells, including prostate cancer cell lines (see, e.g., Batra S, Popper L D, Hartley-Asp B. Prostate. 1991, 19:299).

FIG. 24 shows that cells transfected with a STEAP-2 expression system have enhanced ability to survive exposure to paclitaxel. In order to determine the effect of STEAP-2 on survival, PC3 cells lacking or expressing STEAP-2 were treated with chemotherapeutic agents currently used in the clinic. Effect of treatment was evaluated by measuring cell proliferation using the Alamare blue assay (FIG. 23). While only 5.2% of PC3-neo cells were able to metabolize Alalmare Blue and proliferate in the presence of 5 μM paclitaxel, 44.8% of PC3-STEAP-2 cells survived under the same conditions. These results indicate that expression of STEAP-2 imparts resistance to paclitaxel. These findings have significant in vivo implications, as they indicate that STEAP-2 provides a growth advantage for prostate tumor cells in patients treated with common therapeutic agents.

A more detailed form of these results is shown on FIGS. 25 and 26. Results in these two pages demonstrate the mode of action by which STEAP-2 supports the survival of PC3 cells. In these studies, PC3 cells expressing or lacking STEAP-2 were treated with paclitaxel for 60 hours, and assayed for apoptosis using annexin V conjugated to FITC and propidium iodide staining. In apoptotic cells, the membrane phospholipid phosphatidylserine (PS) is translocated from the inner to the outer leaflet of the membrane, thereby exposing PS to the external cellular environment. PS is recognized by and binds to annexin V, providing scientists with a reliable means of identifying cells undergoing programmed cell death. Staining with propidium iodide identifies dead cells. FIG. 25 show that expression of STEAP-2 inhibits paclitaxel-mediated apoptosis by 45% relative to paclitaxel-treated PC3-neo cells. The protective effect of STEAP-2 is inhibited when STEAP-2 is modified by the presence of Flag at its C-terminus FIG. 26.

The publicly available literature contains several examples of prostate and other cancers that exhibit similar phenotypic characteristics as those observed in PC3 cells that express STEAP-2. In particular, clinical studies have reported transient tumor regression and/or only partial responses in patients treated with paclitaxel. For instance, only around 50% of prostate cancer patients entered in a single agent clinical trial of paclitaxel showed reduced PSA levels when treated with doses of paclitaxel that induced grade 3 and grade 4 toxicity; a much higher level of response would have been expected based on this dose level, thus this data indicates the development of paclitaxel resistance in prostate cancer patients (Beer T M et al., Ann Oncol 2001, 12:1273). A similar phenomenon of reduced responsiveness and progressive tumor recurrence was observed in other studies (see, e.g., Obasaju C, and Hudes G R. Hematol Oncol Clin North Am 2001, 15:525). In addition, inhibition of calcium flux in cells that endogenously express STEAP-2, such as LNCaP cells, induces their cell death (Skryma R et al., J. Physiol. 2000, 527:71).

Thus, STEAP-2 protein is produced not only in the cells tested, but also in unmodified tumor cells or unmodified prostate cells where the presence of mRNA has been shown. The Northern blot data in the specification clearly show that the messenger RNA encoding STEAP-2 is produced in certain prostate and tumor cells. The 3T3 and PC-3 cells, which are themselves tumor cell lines, are clearly able to translate the messenger RNA into protein. Because it has been shown that there is no barrier to translation of the message in cells similar to those tumor and prostate cells in which the mRNA has been shown to be produced, it can properly be concluded that the protein itself can be detected in the unmodified tumor or prostate cells, given the fact that it is shown that mRNA is produced. This conclusion is also supported by the patterns of phenotypic changes seen in cells specifically modified to express STEAP-2, these changes comport with changes seen in cancer cells. Based on the above data, it is scientifically concluded that cells and tissues which produce mRNA encoding STEAP-2 also produce the protein itself.

Example 46 Identification and Confirmation of Potential Signal Transduction Pathways

Many mammalian proteins have been reported to interact with signaling molecules and to participate in regulating signaling pathways (J Neurochem. 2001; 76:217-223. Using immunoprecipitation and Western blotting techniques, proteins are identified that associate with 98P4B6 and mediate signaling events. Several pathways known to play a role in cancer biology can be regulated by 98P4B6, including phospholipid pathways such as PI3K, AKT, etc, adhesion and migration pathways, including FAK, Rho, Rac-1, etc, as well as mitogenic survival cascades such as ERK, p38, etc (Cell Growth Differ. 2000, 11:279; J Biol Chem. 1999, 274:801; Oncogene. 2000, 19:3003, J. Cell Biol. 1997, 138:913.).

To confirm that 98P4B6 directly or indirectly activates known signal transduction pathways in cells, luciferase (luc) based transcriptional reporter assays are carried out in cells expressing individual genes. These transcriptional reporters contain consensus-binding sites for known transcription factors that lie downstream of well-characterized signal transduction pathways. The reporters and examples of these associated transcription factors, signal transduction pathways, and activation stimuli are listed below.

-   -   1. NFkB-luc, NFkB/Rel; lk-kinase/SAPK; growth/apoptosis/stress     -   2. SRE-luc, SRFftCF/ELK1; MAPK/SAPK; growth/differentiation     -   3. AP-1-luc, FOS/JUN; MAPK/SAPK/PKC; growth/apoptosis/stress     -   4. ARE-luc, androgen receptor; steroids/MAPK;         growth/differentiation/apoptosis     -   5. p53-luc, p53; SAPK; growth/differentiation/apoptosis     -   6. CRE-luc, CREB/ATF2; PKA/p38; growth/apoptosis/stress

Gene-mediated effects can be assayed in cells showing mRNA expression. Luciferase reporter plasmids can be introduced by lipid-mediated transfection (TFX-50, Promega). Luciferase activity, an indicator of relative transcriptional activity, is measured by incubation of cell extracts with luciferin substrate and luminescence of the reaction is monitored in a luminometer.

Signaling pathways activated by 98P4B6 are mapped and used for the identification and validation of therapeutic targets. When 98P4B6 is involved in cell signaling, it is used as target for diagnostic, prognostic, preventative and/or therapeutic purposes.

Example 47 98P4B6 Functions as a Proton or Small Molecule Transporter

Sequence and homology analysis of 98P4B6 indicate that the 98P4B6 may function as a transporter. To confirm that STEAP-1 functions as an ion channel, FACS analysis and fluorescent microscopy techniques are used (Gergely L, et al., Clin Diagn Lab Immunol. 1997; 4:70; Skryma R, et al., J. Physiol. 2000, 527: 71). Using FACS analysis and commercially available indicators (Molecular Probes), parental cells and cells expressing 98P4B6 are compared for their ability to transport electrons, sodium, calcium; as well as other small molecules in cancer and normal cell lines. For example, PC3 and PC3-98P4B6 cells were loaded with calcium responsive indicators Fluo4 and Fura red, incubated in the presence or absence of calcium and lipophosphatidic acid (LPA), and analyzed by flow cytometry. Ion flux represents an important mechanism by which cancer cells are regulated. This is particularly true in the case of caldum, as calcium channel inhibitors have been reported to induce the death of certain cancer cells, including prostate cancer cell lines (Batra S, Popper L D, Hartley-Asp B. Prostate. 1991, 19: 299). Similar studies are conducted using sodium, potassium, pH, etc indicators.

Due to its homology to an oxidoreductase, 98P4B6 can participate in imparting drug resistance by mobilizing and transporting small molecules. The effect of 98P4B6 on small molecule transport is investigated using a modified MDR assay. Control and 98P4B6 expressing cells are loaded with a fluorescent small molecule such as calcein AM. Extrusion of calcein from the cell is measured by examining the supernatants for fluorescent compound. MDR-like activity is confirmed using MDR inhibitors.

When 98P4B6 functions as a transporter, it is used as target for diagnostic, prognostic, preventative and/or therapeutic purposes.

Example 48 Involvement in Tumor Progression

The 98P4B6 gene can contribute to the growth of cancer cells. The role of 98P4B6 in tumor growth is confirmed in a variety of primary and transfected cell lines including prostate as well as NIH 3T3 cells engineered to stably express 98P4B6. Parental cells lacking 98P4B6 and cells expressing 98P4B6 are evaluated for cell growth using a well-documented proliferation assay (Fraser S P, Grimes J A, Djamgoz M B. Prostate. 2000; 44:61, Johnson D E, Ochieng J, Evans S L. Anticancer Drugs. 1996, 7:288).

To confirm the role of 98P4B6 in the transformation process, its effect in colony forming assays is investigated. Parental NIH-3T3 cells lacking 98P4B6 are compared to NIH-3T3 cells expressing 98P4B6, using a soft agar assay under stringent and more permissive conditions (Song Z. et al. Cancer Res. 2000; 60:6730).

To confirm the role of 98P4B6 in invasion and metastasis of cancer cells, a well-established assay is used, e.g., a Transwell Insert System assay (Becton Dickinson) (Cancer Res. 1999; 59:6010). Control cells, including prostate and fibroblast cell lines lacking 98P4B6 are compared to cells expressing 98P4B6. Cells are loaded with the fluorescent dye, calcein, and plated in the top well of the Transwell insert coated with a basement membrane analog. Invasion is determined by fluorescence of cells in the lower chamber relative to the fluorescence of the entire cell population.

98P4B6 can also play a role in cell cycle and apoptosis. Parental cells and cells expressing 98P4B6 are compared for differences in cell cycle regulation using a well-established BrdU assay (Abdel-Malek Z A. J Cell Physiol. 1988, 136:247). In short, cells are grown under both optimal (full serum) and limiting (low serum) conditions are labeled with BrdU and stained with anti-BrdU Ab and propidium iodide. Cells are analyzed for entry into the G1, S, and G2M phases of the cell cycle. Alternatively, the effect of stress on apoptosis is evaluated in control parental cells and cells expressing 98P4B6, including normal and tumor prostate cells. Engineered and parental cells are treated with various chemotherapeutic agents, such as etoposide, flutamide, etc, and protein synthesis inhibitors, such as cycloheximide. Cells are stained with annexin V-FITC and cell death is measured by FACS analysis. The modulation of cell death by 98P4B6 can play a critical role in regulating tumor progression and tumor load.

When 98P4B6 plays a role in cell growth, transformation, invasion or apoptosis, it is used as a target for diagnostic, prognostic, preventative and/or therapeutic purposes.

Example 49 Involvement in Angiogenesis

Angiogenesis or new capillary blood vessel formation is necessary for tumor growth (Hanahan D, Folkman J. Cell. 1996, 86:353; Folkman J. Endocrinology. 1998 139:441). Based on the effect of phsophodieseterase inhibitors on endothelial cells, 98P4B6 plays a role in angiogenesis (DeFouw L et al., Microvasc Res 2001, 62:263). Several assays have been developed to measure angiogenesis in vitro and in vivo, such as the tissue culture assays endothelial cell tube formation and endothelial cell proliferation. Using these assays as well as in vitro neo-vascularization, the role of 98P4B6 in angiogenesis, enhancement or inhibition, is confirmed.

For example, endothelial cells engineered to express 98P4B6 are evaluated using tube formation and proliferation assays. The effect of 98P4B6 is also confirmed in animal models in vivo. For example, cells either expressing or lacking 98P4B6 are implanted subcutaneously in immunocompromised mice. Endothelial cell migration and angiogenesis are evaluated 5-15 days later using immunohistochemistry techniques. 98P4B6 affects angiogenesis, and it is used as a target for diagnostic, prognostic, preventative and/or therapeutic purposes.

Example 50 Regulation of Transcription

The localization of 98P4B6 and its similarity to hydrolases as well as its Ets motif (v.7) indicate that 98P4B6 is effectively used as a modulator of the transcriptional regulation of eukaryotic genes. Regulation of gene expression is confirmed, e.g., by studying gene expression in cells expressing or lacking 98P4B6. For this purpose, two types of experiments are performed.

In the first set of experiments, RNA from parental and 98P4B6-expressing cells are extracted and hybridized to commercially available gene arrays (Clontech) (Smid-Koopman E et al. Br J Cancer. 2000. 83:246). Resting cells as well as cells treated with FBS or androgen are compared. Differentially expressed genes are identified in accordance with procedures known in the art. The differentially expressed genes are then mapped to biological pathways (Chen K et al. Thyroid. 2001. 11:41.).

In the second set of experiments, specific transcriptional pathway activation is evaluated using commercially available (Stratagene) luciferase reporter constructs including: NFkB-luc, SRE-luc, ELK1-luc, ARE-luc, p53-luc, and CRE-luc. These transcriptional reporters contain consensus binding sites for known transcription factors that lie downstream of well-characterized signal transduction pathways, and represent a good tool to ascertain pathway activation and screen for positive and negative modulators of pathway activation.

Thus, 98P4B6 plays a role in gene regulation. When 98P4B6 is involved in gene regulation it is used as a target for diagnostic, prognostic, preventative and/or therapeutic purposes.

Example 51 Protein-Protein Association

Several 6TM proteins have been shown to interact with other proteins, thereby regulating signal transduction, gene transcription, transformation, and cell adhesion. Using immunoprecipitation techniques as well as two yeast hybrid systems, proteins are identified that associate with 98P4B6. Immunoprecipitates from cells expressing 98P4B6 and cells lacking 98P4B6 are compared for specific protein-protein associations.

Studies are performed to confirm the extent of association of 98P4B6 with effector molecules, such as nuclear proteins, transcription factors, kinases, phosphates etc. Studies comparing 98P4B6 positive and 98P4B6 negative cells as well as studies comparing unstimulated/resting cells and cells treated with epithelial cell activators, such as cytokines, growth factors, androgen and anti-integrin Ab reveal unique interactions.

In addition, protein-protein interactions are confirmed using two yeast hybrid methodology (Curr Opin Chem. Biol. 1999, 3:64). A vector carrying a library of proteins fused to the activation domain of a transcription factor is introduced into yeast expressing a 98P4B6-DNA-binding domain fusion protein and a reporter construct. Protein-protein interaction is detected by colorimetric reporter activity. Specific association with effector molecules and transcription factors directs one of skill to the mode of action of 98P4B6, and thus identifies therapeutic, prognostic, preventative and/or diagnostic targets for cancer. This and similar assays are also used to identify and screen for small molecules that interact with 98P4B6.

Thus it is found that 98P4B6 associates with proteins and small molecules. Accordingly, 98P4B6 and these proteins and small molecules are used for diagnostic, prognostic, preventative and/or therapeutic purposes.

Throughout this application, various website data content, publications, patent applications and patents are referenced. (Websites are referenced by their Uniform Resource Locator, or URL, addresses on the World Wide Web.) The disclosures of each of these references are hereby incorporated by reference herein in their entireties.

The present invention is not to be limited in scope by the embodiments disclosed herein, which are intended as single illustrations of individual aspects of the invention, and any that are functionally equivalent are within the scope of the invention. Various modifications to the models and methods of the invention, in addition to those described herein, will become apparent to those skilled in the art from the foregoing description and teachings, and are similarly intended to fall within the scope of the invention. Such modifications or other embodiments can be practiced without departing from the true scope and spirit of the invention.

Tables:

TABLE I Tissues that Express 98P4B6: a. Malignant Tissues a Bladder b. Breast c. Cervix d. Colon e. Kidney f. Lung g. Ovary h. Pancreas i. Prostate j. Stomach k. Uterus

TABLE II Amino Acid Abbreviations SINGLE LETTER THREE LETTER FULL NAME F Phe phenylalanine L Leu leucine S Ser serine Y Tyr tyrosine C Cys cysteine W Trp tryptophan P Pro proline H His histidine Q Gln glutamine R Arg arginine I Ile isoleucine M Met methionine T Thr threonine N Asn asparagine K Lys lysine V Val valine A Ala alanine D Asp aspartic acid E Glu glutamic acid G Gly glycine

TABLE III Amino Acid Substitution Matrix Adapted from the GCG Software 9.0 BLOSUM62 amino acid substitution matrix (block substitution matrix). The higher the value, the more likely a substitution is found in related, natural proteins. A C D E F G H I K L M N P Q R S T V W Y . 4 0 −2 −1 −2 0 −2 −1 −1 −1 −1 −2 −1 −1 −1 1 0 0 −3 −2 A 9 −3 −4 −2 −3 −3 −1 −3 −1 −1 −3 −3 −3 −3 −1 −1 −1 −2 −2 C 6 2 −3 −1 −1 −3 −1 −4 −3 1 −1 0 −2 0 −1 −3 −4 −3 D 5 −3 −2 0 −3 1 −3 −2 0 −1 2 0 0 −1 −2 −3 −2 E 6 −3 −1 0 −3 0 0 −3 −4 −3 −3 −2 −2 −1 1 3 F 6 −2 −4 −2 −4 −3 0 −2 −2 −2 0 −2 −3 −2 −3 G 8 −3 −1 −3 −2 1 −2 0 0 −1 −2 −3 −2 2 H 4 −3 2 1 −3 −3 −3 −3 −2 −1 3 −3 −1 I 5 −2 −1 0 −1 1 2 0 −1 −2 −3 −2 K 4 2 −3 −3 −2 −2 −2 −1 1 −2 −1 L 5 −2 −2 0 −1 −1 −1 1 −1 −1 M 6 −2 0 0 1 0 −3 −4 −2 N 7 −1 −2 −1 −1 −2 −4 −3 P 5 1 0 −1 −2 −2 −1 Q 5 −1 −1 −3 −3 −2 R 4 1 −2 −3 −2 S 5 0 −2 −2 T 4 −3 −1 V 11 2 W 7 Y

TABLE IV HLA Class I/II Motifs/Supermotifs TABLE IV (A): HLA Class I Supermotifs/Motifs POSITION POSITION POSITION C Terminus (Primary 2 (Primary Anchor) 3 (Primary Anchor) Anchor) SUPERMOTIF A1 TI LVMS FWY A2 LIVM ATQ IV MATL A3 VSMA TLI RK A24 YF WIVLMT FI YWLM B7 P VILF MWYA B27 RHK FYL WMIVA B44 E D FWYLIMVA B58 ATS FWY LIVMA B62 QL IVMP FWY MIVLA MOTIFS A1 TSM Y A1 DE AS Y A2.1 LM VQIAT V LIMAT A3 LMVISATF CGD KYR HFA A11 VTMLISAGN CDF K RYH A24 YF WM FLIW A*3101 MVT ALIS R K A*3301 MVALF IST RK A*6801 AVT MSLI RK B*0702 P LMF WYAIV B*3501 P LMFWY IVA B51 P LIVF WYAM B*5301 P IMFWY ALV B*5401 P ATIV LMFWY Bolded residues are preferred, italicized residues are less preferred: A peptide is considered motif-bearing if it has primary anchors at each primary anchor position for a motif or supermotif as specified in the above table. TABLE IV (B): HLA Class II Supermotif 1 6 9 W, F, Y, V, .I, L A, V, I, L, P, C, S, T A, V, I, L, C, S, T, M, Y TABLE IV (C): HLA Class II Motifs MOTIFS 1° anchor 1 2 3 4 5 1° anchor 6 7 8 9 DR4 preferred FMYLIVW M T I VSTCPALIM MH MH deleterious W R WDE DR1 preferred MFLIVWY PAMQ VMATSPLIC M AVM deleterious C CH FD CWD GDE D DR7 preferred MFLIVWY M W A IVMSACTPL M IV deleterious C G GRD N G DR3 MOTIFS 1° anchor 1 2 3 1° anchor 4 5 1° anchor 6 Motif a preferred LIVMFY D Motif b preferred LIVMFAY DNQEST KRH DR Supermotif MFLIVWY VMSTACPLI Italicized residues indicate less preferred or “tolerated” residues TABLE IV (D): HLA Class I Supermotifs SUPER- POSITION: MOTIFS 1 2 3 4 5 6 7 8 C-terminus A1 1° Anchor 1° Anchor TILVMS FWY A2 1° Anchor 1° Anchor LIVMATQ LIVMAT A3 Preferred 1° Anchor YFW YFW YFW P 1° Anchor VSMATLI (4/5) (3/5) (4/5) (4/5) RK deleterious DE (3/5); DE P (5/5) (4/5) A24 1° Anchor 1° Anchor YFWIVLMT FIYWLM B7 Preferred FWY (5/5) 1° Anchor FWY FWY 1° Anchor LIVM (3/5) P (4/5) (3/5) VILFMWYA deleterious DE (3/5); DE G QN DE P (5/5); (3/5) (4/5) (4/5) (4/5) G (4/5); A (3/5); QN (3/5) B27 1° Anchor 1° Anchor RHK FYLWMIVA B44 1° Anchor 1° Anchor ED FWYLIMVA B58 1° Anchor 1° Anchor ATS FWYLIVMA B62 1° Anchor 1° Anchor QLIVMP FWYMIVLA Italicized residues indicate less preferred or “tolerated” residues TABLE IV (E): HLA Class I Motifs POSITION 1 2 3 4 5 A1 preferred GFYW 1° Anchor DEA YFW 9-mer STM deleterious DE RHKLIVMP A G A1 preferred GRHK ASTCLIVM 1° Anchor GSTC 9-mer DEAS deleterious A RHKDEPYFW DE PQN A1 preferred YFW 1° Anchor DEAQN A YFWQN 10- STM mer deleterious GP RHKGLIVM DE RHK A1 preferred YFW STCLIVM 1° Anchor A YFW 10- DEAS mer deleterious RHK RHKDEPYFW P A2.1 preferred YFW 1° Anchor YFW STC YFW 9-mer LMIVQAT deleterious DEP DERKH A2.1 preferred AYFW 1° Anchor LVIM G 10- LMIVQAT mer deleterious DEP DE RKHA P A3 preferred RHK 1° Anchor YFW PRHKYFW A LMVISATFCGD deleterious DEP DE A11 preferred A 1° Anchor YFW YFW A VTLMISAGNCDF deleterious DEP A24 preferred YFWRHK 1° Anchor STC 9-mer YFWM deleterious DEG DE G QNP A24 Preferred 1° Anchor P YFWP 10- YFWM mer Deleterious GDE QN RHK A3101 Preferred RHK 1° Anchor YFW P MVTALIS Deleterious DEP DE ADE A3301 Preferred 1° Anchor YFW MVALFIST Deleterious GP DE A6801 Preferred YFWSTC 1° Anchor YFWLIVM AVTMSLI deleterious GP DEG RHK B0702 Preferred RHKFWY 1° Anchor RHK RHK P deleterious DEQNP DEP DE DE B3501 Preferred FWYLIVM 1° Anchor FWY P A1 preferred GFYW 1° Anchor DEA YFW 9-mer STM deleterious DE RHKLIVMP A G A1 preferred GRHK ASTCLIVM 1° Anchor GSTC 9-mer DEAS deleterious A RHKDEPYFW DE PQN deleterious AGP G B51 Preferred LIVMFWY 1° Anchor FWY STC FWY P deleterious AGPDER DE HKSTC B5301 preferred LIVMFWY 1° Anchor FWY STC FWY P deleterious AGPQN B5401 preferred FWY 1° Anchor FWYLIVM LIVM P deleterious GPQNDE GDESTC RHKDE POSITION 9 or 6 7 8 C-terminus C-terminus A1 preferred P DEQN YFW 1° Anchor 9-mer Y deleterious A A1 preferred ASTC LIVM DE 1° Anchor 9-mer Y deleterious RHK PG GP A1 preferred PASTC GDE P 1° Anchor 10- Y mer deleterious QNA RHKYFW RHK A A1 preferred PG G YFW 1° Anchor 10- Y mer deleterious G PRHK QN A2.1 preferred A P 1° Anchor 9-mer VLIMAT deleterious RKH DERKH A2.1 preferred G FYWL 1° Anchor 10- VIM VLIMAT mer deleterious RKH DERKH RKH A3 preferred YFW P 1° Anchor KYRHFA deleterious A11 preferred YFW YFW P 1° Anchor KRYH deleterious A G A24 preferred YFW YFW 1° Anchor 9-mer FLIW deleterious DERHK G AQN A24 Preferred P 1° Anchor 10- FLIW mer Deleterious DE A QN DEA A3101 Preferred YFW YFW AP 1° Anchor RK Deleterious DE DE DE A3301 Preferred AYFW 1° Anchor RK Deleterious A6801 Preferred YFW P 1° Anchor RK deleterious A B0702 Preferred RHK RHK PA 1° Anchor LMFWYAIV deleterious GDE QN DE B3501 Preferred FWY 1° Anchor LMFWYIVA A1 preferred P DEQN YFW 1° Anchor 9-mer Y deleterious A A1 preferred ASTC LIVM DE 1° Anchor 9-mer Y deleterious RHK PG GP deleterious G B51 Preferred G FWY 1° Anchor LIVFWYAM deleterious G DEQN GDE B5301 preferred LIVMFWY FWY 1° Anchor IMFWYALV deleterious G RHKQN DE B5401 preferred ALIVM FWYAP 1° Anchor ATIVLMFWY deleterious DE QNDGE DE TABLE IV (F): Summary of HLA-supertypes Overall phenotypic frequencies of HLA-supertypes in different ethnic populations Specificity Phenotypic frequency Supertype Position 2 C-Terminus Caucasian N.A. Black Japanese Chinese Hispanic Average B7 P AILMVFWY 43.2 55.1 57.1 43.0 49.3 49.5 A3 AILMVST RK 37.5 42.1 45.8 52.7 43.1 44.2 A2 AILMVT AILMVT 45.8 39.0 42.4 45.9 43.0 42.2 A24 YF (WIVLMT) FI (YWLM) 23.9 38.9 58.6 40.1 38.3 40.0 B44 E (D) FWYLIMVA 43.0 21.2 42.9 39.1 39.0 37.0 A1 TI (LVMS) FWY 47.1 16.1 21.8 14.7 26.3 25.2 B27 RHK FYL (WMI) 28.4 26.1 13.3 13.9 35.3 23.4 B62 QL (IVMP) FWY (MIV) 12.6  4.8 36.5 25.4 11.1 18.1 B58 ATS FWY (LIV) 10.0 25.1  1.6  9.0  5.9 10.3 TABLE IV (G): Calculated population coverage afforded by different HLA-supertype combinations Phenotypic frequency HLA-supertypes Caucasian N.A Blacks Japanese Chinese Hispanic Average A2, A3, and B7 83.0 86.1  87.5 88.4 86.3 86.2 A2, A3, B7, A24, B44 and 99.5 98.1 100.0 99.5 99.4 99.3 A1 A2, A3, B7, A24, B44, 99.9 99.6 100.0 99.8 99.9 99.8 A1, B27, B62, and B58 Motifs indicate the residues defining supertype specificites. The motifs incorporate residues determined on the basis of published data to be recognized by multiple alleles within the supertype. Residues within brackets are additional residues also predicted to be tolerated by multiple alleles within the supertype.

TABLE V Frequently Occurring Motifs avrg. % Name identity Description Potential Function zf-C2H2 34% Zinc finger, C2H2 type Nucleic acid-binding protein functions as transcription factor, nuclear location probable cytochrome_b_N 68% Cytochrome b(N- membrane bound oxidase, generate terminal)/b6/petB superoxide Ig 19% Immunoglobulin domain domains are one hundred amino acids long and include a conserved intradomain disulfide bond. WD40 18% WD domain, G-beta repeat tandem repeats of about 40 residues, each containing a Trp-Asp motif. Function in signal transduction and protein interaction PDZ 23% PDZ domain may function in targeting signaling molecules to sub-membranous sites LRR 28% Leucine Rich Repeat short sequence motifs involved in protein-protein interactions Pkinase 23% Protein kinase domain conserved catalytic core common to both serine/threonine and tyrosine protein kinases containing an ATP binding site and a catalytic site PH 16% PH domain pleckstrin homology involved in intracellular signaling or as constituents of the cytoskeleton EGF 34% EGF-like domain 30-40 amino-acid long found in the extracellular domain of membrane- bound proteins or in secreted proteins Rvt 49% Reverse transcriptase (RNA-dependent DNA polymerase) Ank 25% Ank repeat Cytoplasmic protein, associates integral membrane proteins to the cytoskeleton Oxidored_q1 32% NADH- membrane associated. Involved in Ubiquinone/plastoquinone proton translocation across the (complex I), various chains membrane Efhand 24% EF hand calcium-binding domain, consists of a12 residue loop flanked on both sides by a 12 residue alpha-helical domain Rvp 79% Retroviral aspartyl Aspartyl or acid proteases, centered on protease a catalytic aspartyl residue Collagen 42% Collagen triple helix repeat extracellular structural proteins involved (20 copies) in formation of connective tissue. The sequence consists of the G-X-Y and the polypeptide chains forms a triple helix. Fn3 20% Fibronectin type III domain Located in the extracellular ligand- binding region of receptors and is about 200 amino acid residues long with two pairs of cysteines involved in disulfide bonds 7tm_1 19% 7 transmembrane receptor seven hydrophobic transmembrane (rhodopsin family) regions, with the N-terminus located extracellularly while the C-terminus is cytoplasmic. Signal through G proteins

TABLE VI Motifs and Post-translational Modifications of 98P4B6 cAMP- and cGMP-dependent protein kinase phosphorylation site. 176-179 RKET (SEQ ID NO: 114) Protein kinase C phosphorylation site. 235-237 SVK Casein kinase II phosphorylation site.   9-12 SATD (SEQ ID NO: 115)  50-53 TVME (SEQ ID NO: 116) 130-133 SCTD (SEQ ID NO: 117) 172-175 SPEE (SEQ ID NO: 118) N-myristoylation site.  14-19 GLSIST (SEQ ID NO: 119) G-protein coupled receptors family 1 signature.  52-68 MESSVLLAMAFDRFVAV (SEQ ID NO: 120)

TABLE VII Search Peptides v.1 aa1-454 (SEQ ID NO: 121) 9-mers, 10-mers and 15-mers MESISMMGSP KSLSETCLPN GINGIKDARK VTVGVIGSGD FAKSLTIRLI RCGYHVVIGS RNPKFASEFF PHVVDVTHHE DALTKTNIIF VAIHREHYTS LWDLRHLLVG KILIDVSNNM RINQYPESNA EYLASLFPDS LIVKGFNVVS AWALQLGPKD ASRQVYICSN NIQARQQVIE LARQLNFIPI DLGSLSSARE IENLPLRLFT LWRGPVVVAI SLATFFFLYS FVRDVIHPYA RNQQSDFYKI PIEIVNKTLP IVAITLLSLV YLAGLLAAAY QLYYGTKYRR FPPWLETWLQ CRKQLGLLSF FFAMVHVAYS LCLPMRRSER YLFLNMAYQQ VHANIENSWN EEEVWRIEMY ISFGIMSLGL LSLLAVTSIP SVSNALNWRE FSFIQSTLGY VALLISTFHV LIYGWKRAFE EEYYRFYTPP NFVLALVLPS IVILDLLQLC RYPD v.2 aa1-45 (SEQ ID NO: 122) 9-mers, 10-mers, 15-mers SGSPGLQALSL SLSSGFTPFS CLSLPSSWDY RCPPPCPADF FLYF v.5, (one aa diff at 211 and different c-terminal) Part A 9-mers: aa203-219 (SEQ ID NO: 123) NLPLRLFTFWRGPVVVA 10-mers: aa202-220 (SEQ ID NO: 124) ENLPLRLFTFWRGPVVVAI 15-mers: aa197-225 (SEQ ID NO: 125) SAREIENLPLRLFTFWRGPVVVAISLATF Part B 9-mers: aa388-419 (SEQ ID NO: 126) WREFSFIQIFCSFADTQTELELEFVFLLTLLL 10-mers: aa387-419 (SEQ ID NO: 127) NWREFSFIQIFCSFADTQTELELEFVFLLTLLL 15-mers: aa382-419 (SEQ ID NO: 128) VSNALNWREFSFIQIFCSFADTQTELELEFVFLLTLLL v.6, (different from our original in 445-490) 9-mers; aa447-490 (SEQ ID NO: 129) VLPSIVILGKIILFLPCISRKLKRIKKGWEKSQFLEEGIGGTIPHVSPER VTVM 10-mers: aa446-490 (SEQ ID NO: 130) LVLPSIVILGKIILFLPCISRKLKRIKKGWEKSQFLEEGIGGTIPHVSPE RVTVM 15-mers: aa441-490 (SEQ ID NO: 131) NFVLALVLPSIVILGKIILFLPCISRKLKRIKKGWEKSQFLEEGIGGTIP HVSPERVTVM v.7, (deleting our original 340-394, 392-576 is different) Part A 9-mers: aa334-350 (SEQ ID NO: 132) FLNMAYQQSTLGYVALL 10-mers: aa333-351 (SEQ ID NO: 133) LFLNNAYQQSTLGYVALLI 15-mers: aa328-355 (SEQ ID NO: 134) RSERYLFLNMAYQQSTLGYVALLISTFHV Part B 9-mers: aa384-576 (SEQ ID NO: 135) PSIVILDLSVEVLASPAAAWKCLGANILRGGLSEIVLPIEWQQDRKIPPL STPPPPA MWTEEAGATAEAQESGIRNKSSSSSQIPVVGVVTEDDEAQDSIDPPESPD RALKAANSWRNPVLPHTNGVGPLWEFLLRLLKSQAASGTLSLAFTSWSLG EFLGSGTWMKLETIILSKLTQEQKSKHCMF SLISGS 10-mers: aa383-576 (SEQ ID NO: 136) LPSIVILDLSVEVLASPAAAWKCLGANILRGGLSEIVLPIEWQQDRKIPP LSTPPPPA MWTEEAGATAEAQESGIRNKSSSSSQIPVVGVVTEDDEAQDSIDPPESPD RALKAANSWRNPVLPHTNGVGPLWEFLLRLLKSQAASGTLSLAFTSWSLG EFLGSGTWMK LETIILSKLT QEQKSKHCMF SLISGS 15-mers: aa378-576 (SEQ ID NO: 137) VLALVLPSIVILDLSVEVLASPAAAWKCLGANILRGGLSEIVLPIEWQQD RKIPPLSTPPPPAMWTEEAGATAEAQESGIRNKSSSSSQIPVVGVVTEDD EAQDSIDPPESPDRALKAANSWRNPVLPHTNGVGPLWEFLLRLLKSQAAS GTLSLAFTSWSLG EFLGSGTWMK LETIILSKLT QEQKSKHCMF SLISGS v.8, SNP variant of v.6, one aa different at 475 9-mers: aa466-482 (SEQ ID NO: 138) KSQFLEEGMGGTIPHVS 10-mers: aa465-483 (SEQ ID NO: 139) EKSQFLEEGMGGTIPHVSP 15-mers: aa460-489 (SEQ ID NO: 140) IKKGWEKSQFLEEGMGGTIPHVSPERVTV V13 9-mers: aa9-25 (SEQ ID NO: 141) SPKSLSETFLPNGINGI 10-mers: aa8-26 (SEQ ID NO: 142) GSPKSLSETFLPNGINGIK 15-mers: aa3-31 (SEQ ID NO: 143) SISMMGSPKSLSETFLPNGINGIKDARKV v.14 9-mers: aa203-219 (SEQ ID NO: 144) NLPLRLFTFWRGPVVVA 10-mers: aa202-220 (SEQ ID NO: 145) ENLPLRLFTFWRGPVVVAI 15-mers: aa197-225 (SEQ ID NO: 146) SAREIENLPLRLFTFWRGPVVVAISLATF V.21 9-mers 557-572 (SEQ ID NO: 147) SKLTQEQKTKHCMFSLI 10-mers 556-573 (SEQ ID NO: 148) LSKLTQEQKTKHCMFSLIS 15-mers 551-576 (SEQ ID NO: 149) LETIILSKLTQEQKTKHCMFSLISGS V.25 9-mers aa 447-463 (SEQ ID NO: 150) ILFLPCISQKLKRIKKG 10-mers aa 446-464 (SEQ ID NO: 151) IILFLPCISQKLKRIKKGW 15-mers aa440-468 (SEQ ID NO: 152) VILGKIILFLPCISQKLKRIKKGWEKSQF

TABLE VIII-V1 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight Start Subsequence Score 443 ILDLLQLCR 25.000 129 NAEYLASLF 9.000 294 WLETWLQCR 9.000 113 LIDVSNNMR 5.000 200 EIENLPLRL 4.500 244 QSDFYKIPI 3.750 405 ISTFHVLIY 3.750 13 LSETCLPNG 2.700 221 SLATFFFLY 2.500 263 AITLLSLVY 2.500 276 LAAAYQLYY 2.500 419 FEEEYYRFY 2.250 155 QLGPKDASR 2.000 66 ASEFFPHVV 1.350 272 LAGLLAAAY 1.000 35 VIGSGDFAK 1.000 178 VIELARQLN 0.900 356 RIEMYISFG 0.900 418 AFEEEYYRF 0.900 319 YSLCLPMRR 0.750 43 KSLTIRLIR 0.750 327 RSERYLFLN 0.675 427 YTPPNFVLA 0.500 304 QLGLLSFFF 0.500 257 KTLPIVAIT 0.500 135 SLFPDSLIV 0.500 223 ATFFFLYSF 0.500 275 LLAAAYQLY 0.500 385 ALNWREFSF 0.500 219 AISLATFFF 0.500 16 TCLPNGING 0.500 90 FVAIHREHY 0.500 87 NIIFVAIHR 0.500 249 KIPIEIVNK 0.400 137 FPDSLIVKG 0.250 189 PIDLGSLSS 0.250 241 RNQQSDFYK 0.250 351 EEEVWRIEM 0.225 349 WNEEEVWRI 0.225 125 YPESNAEYL 0.225 420 EEEYYRFYT 0.225 388 WREFSFIQS 0.225 198 AREIENLPL 0.225 57 VIGSRNPKF 0.200 56 VVIGSRNPK 0.200 217 VVAISLATF 0.200 3 SISMMGSPK 0.200 417 RAFEEEYYR 0.200 436 LVLPSIVIL 0.200 377 TSIPSVSNA 0.150 158 PKDASRQVY 0.125 101 KWDLRHLLV 0.125 117 SNNMRINQY 0.125 392 SFIQSTLGY 0.125 202 ENLPLRLFT 0.125 330 RYLFLNMAY 0.125 38 SGDFAKSLT 0.125 98 YTSLWDLRH 0.125 406 STFHVLIYG 0.125 218 VAISLATFF 0.100 167 ICSNNIQAR 0.100 400 YVALLISTF 0.100 235 VIHPYARNQ 0.100 381 SVSNALNWR 0.100 22 INGIKDARK 0.100 21 GINGIKDAR 0.100 281 QLYYGTKYR 0.100 322 CLPMRRSER 0.100 411 LIYGWKRAF 0.100 191 DLGSLSSAR 0.100 409 HVLIYGWKR 0.100 344 NIENSWNEE 0.090 251 PIEIVNKTL 0.090 308 LSFFFAMVH 0.075 195 LSSAREIEN 0.075 116 VSNNMRINQ 0.075 280 YQIYYGTKY 0.075 220 ISLATFFFL 0.075 175 RQQVIELAR 0.075 127 ESNAEYLAS 0.075 432 FVLALVLPS 0.050 12 SLSETCLPN 0.050 106 HLLVGKILI 0.050 311 FFAMVHVAY 0.050 269 LVYLAGLLA 0.050 216 VVVAISLAT 0.050 124 QYPESNAEY 0.050 166 YICSNNIQA 0.050 258 TLPIVAITL 0.050 18 LPNGINGIK 0.050 435 ALVLPSIVI 0.050 25 IKDARKVTV 0.050 73 VVDVTHHED 0.050 222 LATFFFLYS 0.050 184 QLNFIPIDL 0.050 367 SLGLLSLLA 0.050 46 TIRLIRCGY 0.050 306 GLLSFFFAM 0.050 261 IVAITLLSL 0.050 203 NLPLRLFTL 0.050

TABLE VIII-V2 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 23 LSLPSSWDY 7.500 33 CPPPCPADF 0.500 36 PCPADFFLY 0.250 9 LSLSLSSGF 0.150 37 CPADFFLYF 0.125 17 FTPFSCLSL 0.125 24 SLPSSWDYR 0.100 12 SLSSGFTPF 0.100 14 SSGFTPFSC 0.075 5 GLQALSLSL 0.050 7 QALSLSLSS 0.050 13 LSSGFTPFS 0.030 2 GSPGLQALS 0.030 20 FSCLSLPSS 0.030 1 SGSPGLQAL 0.025 32 RCPPPCPAD 0.020 35 PPCPADFFL 0.013 3 SPGLQALSL 0.013 21 SCLSLPSSW 0.010 8 ALSLSLSSG 0.010 10 SLSLSSGFT 0.010 11 LSLSSGFTP 0.007 25 LPSSWDYRC 0.005 16 GFTPFSCLS 0.005 28 SWDYRCPPP 0.005 31 YRCPPPCPA 0.005 15 SGFTPFSCL 0.003 34 PPPCPADFF 0.003 6 LQALSLSLS 0.002 22 CLSLPSSWD 0.001 19 PFSCLSLPS 0.000 18 TPFSCLSLP 0.000 4 PGLQALSLS 0.000 27 SSWDYRCPP 0.000 26 PSSWDYRCP 0.000 29 WDYRCPPPC 0.000 30 DYRCPPPCP 0.000

TABLE VIII-V5A HLA-A1-19mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 NLPLRLFTF 0.500 7 FTFWRGPVV 0.050 3 PLRLFTFWR 0.005 5 RLFTFWRGP 0.001 6 LFTFWRGPV 0.001 4 LRLFTFWRG 0.001 2 LPLRLFTFW 0.000 9 FWRGPVVVA 0.000 8 TFWRGPVVV 0.000

TABLE VIII-V5B HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 21 ELEFVFLLT 4.500 17 QTELELEFV 2.250 19 ELELEFVFL 1.800 1 WREESFIQI 0.225 16 TQTELELEF 0.075 4 FSFIQIFCS 0.075 24 FVFLLTLLL 0.050 13 FADTQTELE 0.050 18 TELELEFVF 0.025 8 QIFCSFADT 0.020 10 FCSFADTQT 0.010 6 FIQIFDSFA 0.010 2 REFSFIQIF 0.005 5 SFIQIFCSF 0.005 15 DTQTELELE 0.003 20 LELEFVFLL 0.003 22 LEFVFLLTL 0.003 14 ADTQTELEL 0.003 3 EFSFIQIFC 0.003 11 CSFADTQTE 0.002 7 IQIFOSFAD 0.001 23 EFVFLLTLL 0.001 12 SFADTQTEL 0.001 9 IFCSFADTQ 0.001

TABLE VIII-V6 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 34 FLEEGIGGT 0.900 12 ILFLPCISR 0.500 6 VILGKIILF 0.500 2 LPSIVILGK 0.250 42 TIPHVSPER 0.200 45 HVSPERVTV 0.200 13 LFLPCISRK 0.100 16 PCISRKLKR 0.050 1 VLPSIVILG 0.050 15 LPCISRKLK 0.050 5 IVILGKIIL 0.050 35 LEEGIGGTI 0.045 41 GTIPHVSPE 0.025 38 GIGGTIPHV 0.020 10 KIILFLPCI 0.020 31 KSQFLEEGI 0.015 46 VSPERVTVM 0.015 37 EGIGGTIPH 0.013 4 SIVILGKII 0.010 14 FLPCISRKL 0.010 11 IILFLPCIS 0.010 19 SRKLKRIKK 0.005 7 ILGKIILFL 0.005 26 KKGWEKSQF 0.005 18 ISRKLKRIK 0.003 33 QFLEEGIGG 0.003 43 IPHVSPERV 0.003 9 GKILLFLPC 0.003 39 IGGTIPHVS 0.003 28 GWEKSQFLE 0.002 3 PSIVILGKI 0.002 32 SQFLEEGIG 0.002 23 KRIKKGWEK 0.001 17 CISRKLKRI 0.001 40 GGTIPHVSP 0.001 30 EKSQFLEEG 0.001 27 KGWEKSQFL 0.000 8 LGKIILFLP 0.000 24 RIKKGWEKS 0.000 21 KLKRIKKGW 0.000 36 EEGIGGTIP 0.000 44 PHVSPERVT 0.000 20 RKLKRIKKG 0.000 25 IKKGWEKSQ 0.000 29 WEKSQFLEE 0.000 22 LKRIKKGWE 0.000

TABLE VIII-V7A HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 5 LSETFLPNG 2.700 4 SLSETFLPN 0.050 7 ETFLPNGIN 0.025 8 TFLPNGING 0.025 9 FLPNGINGI 0.010 3 KSLSETFLP 0.007 1 SPKSLSETF 0.003 6 SETFLPNGI 0.001 2 PKSLSETFL 0.000

TABLE VIII-V7B HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 5 AYQQSTLGY 0.125 9 STLGYVALL 0.050 8 QSTLGYVAL 0.030 1 FLNMAYQQS 0.010 4 MAYQQSTLG 0.010 3 NMAYQQSTL 0.005 7 QQSTLGYVA 0.003 2 LNMAYQQST 0.003 6 YQQSTLGYV 0.002

TABLE VIII-V7C HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 167 KLETIILSK 90.000 59 WTEEAGATA 4.500 13 LASPAAAWK 4.000 69 AQESGIRNK 2.700 38 PIEWQQDRK 1.800 66 TAEAQESGI 0.900 9 SVEVLASPA 0.900 143 ASGTLSLAF 0.750 99 SIDPPESPD 0.500 51 STPPPPAMW 0.500 5 ILDLSVEVL 0.500 21 KCLGANILR 0.500 90 VTEDDEAQD 0.450 50 LSTPPPPAM 0.300 32 LSEIVLPIE 0.270 151 FTSWSLGEF 0.250 156 LGEFLGSGT 0.225 175 KLTQEQKSK 0.200 159 FLGSGTWMK 0.200 177 TQEQKSKHC 0.135 128 GPLWEFLLR 0.125 145 GTLSLAFTS 0.125 52 TPPPPAMWT 0.125 126 GVGPLWEFL 0.100 35 IVLPIEWQQ 0.100 100 IDPPESPDR 0.100 104 ESPDRALKA 0.075 78 SSSSSQIPV 0.075 154 WSLGEFLGS 0.075 131 WEFLLRLLK 0.050 22 CLGANILRG 0.050 68 EAQESGIRN 0.050 184 HCMFSLISG 0.050 7 DLSVEVLAS 0.050 170 TIILSKLTQ 0.050 2 SIVILDLSV 0.050 17 AAAWKCLGA 0.050 141 QAASGTLSL 0.050 123 HTNGVGPLW 0.050 31 GLSEIVLPI 0.050 130 LWEFLLRLL 0.045 173 LSKLTQEQK 0.030 80 SSSQIPVVG 0.030 81 SSQIPVVGV 0.030 79 SSSSQIPVV 0.030 125 NGVGPLWEF 0.025 65 ATAEAQESG 0.025 37 LPIEWQQDR 0.025 92 EDDEAQDSI 0.025 169 ETIILSKLT 0.025 176 LTQEQKSKH 0.025 91 TEDDEAQDS 0.025 102 PPESPDRAL 0.022 103 PESPDRALK 0.020 11 EVLASPAAA 0.020 83 QIPVVGVVT 0.020 4 VILDLSVEV 0.020 12 VLASPAAAW 0.020 42 QQDRKIPPL 0.015 71 ESGIRNKSS 0.015 96 AQDSIDPPE 0.015 14 ASPAAAWKC 0.015 82 SQIPVVGVV 0.015 139 KSQAASGTL 0.015 147 LSLAFTSWS 0.015 29 RGGLSEIVL 0.013 105 SPDRALKAA 0.013 162 SGTWMKLET 0.013 160 LGSGTWMKL 0.013 127 VGPLWEFLL 0.013 146 TLSLAFTSW 0.010 88 GVVTEDDEA 0.010 142 AASGTLSLA 0.010 64 GATAEAQES 0.010 119 PVLPHTNGV 0.010 46 KIPPLSTPP 0.010 62 EAGATAEAQ 0.010 109 ALKAANSWR 0.010 148 SLAFTSWSL 0.010 112 AANSWRNPV 0.010 149 LAFTSWSLG 0.010 34 EIVLPIEWQ 0.010 116 WRNPVLPHT 0.010 24 GANILRGGL 0.010 89 VVTEDDEAQ 0.010 155 SLGEFLGSG 0.010 120 VLPHTNGVG 0.010 181 KSKHCMFSL 0.008 113 ANSWRNPVL 0.005 67 AEAQESGIR 0.005 185 CMFSLISGS 0.005 144 SGTLSLAFT 0.005 93 DDEAQDSID 0.005 60 TEEAGATAE 0.005 8 LSVEVLASP 0.003 183 KHCMFSLIS 0.003 25 ANILRGGLS 0.003 165 WMKLETIIL 0.003 101 DPPESPDRA 0.003 15 SPAAAWKCL 0.003

TABLE IX-V1 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 178 VIELARQLNF 45.000 443 ILDLLQLCRY 25.000 294 WLETWLQCRK 18.000 135 SLFPDSLIVK 10.000 200 EIENLPLRLF 9.000 356 RIEMYISFGI 4.500 220 ISLATFFFLY 3.750 391 FSFIQSTLGY 3.750 76 VTHHEDALTK 2.500 404 LISTFHVLIY 2.500 262 VAITLLSLVY 2.500 275 LLAAAYQLYY 2.500 113 LIDVSNNMRI 2.500 351 EEEVWRIEMY 2.250 418 AFEEEYYRFY 2.250 123 NQYPESNAEY 1.500 13 LSETCLPNGI 1.350 137 FPDSLIVKGF 1.250 427 YTPPNFVLAL 1.250 257 KTLPIVAITL 1.250 271 YLAGLLAAAY 1.000 34 GVIGSGDFAK 1.000 321 LCLMMRRSER 1.000 198 AREIENLPLR 0.900 116 VSNNMRINQY 0.750 327 RSERYLFLNM 0.675 38 SGDFAKSLTI 0.625 384 NALNWREFSF 0.500 218 VAISLATFFF 0.500 274 GLLAAAYQLY 0.500 81 DALTKTNIIF 0.500 322 CLPMRRSERY 0.500 73 VVDVTHHEDA 0.500 232 VRDVIHPYAR 0.500 442 VILDLLQLCR 0.500 125 YPESNAEYLA 0.450 129 NAEYLASLFP 0.450 21 GINGIKDARK 0.400 2 ESISMMGSPK 0.300 66 ASEFFPHVVD 0.270 419 FEEEYYRFYT 0.225 350 NEEEFWRIEM 0.225 222 LATFFFLYSF 0.200 56 VVIGSRNPKF 0.200 281 QLYYGTKYRR 0.200 55 HVVIGSRNPK 0.200 278 AAYQLYYGTK 0.200 417 RAFEEEYYRF 0.200 216 VVVAISLATF 0.200 248 YKIPIEIVNK 0.200 317 VAYSLCLPMR 0.200 17 CLPNGINGIK 0.200 244 QSDFYKIPIE 0.150 377 TSIPSVSNAL 0.150 382 VSNALNWREF 0.150 202 ENLPLRLFTL 0.125 101 LWDLRHLLVG 0.125 329 ERYLFLNMAY 0.125 15 ETCLPNGING 0.125 396 STLGYVALLI 0.125 45 LTIRLIRCGY 0.125 86 TNIIFVAIHR 0.125 32 TVGVIGSGDF 0.100 235 VIHPYARNQQ 0.100 410 VLIYGWKRAF 0.100 112 ILIDVSNNMR 0.100 166 YICSNNIQAR 0.100 16 TCLPNGINGI 0.100 217 VVAISLATFF 0.100 155 QLGPKDASRQ 0.100 344 NIENSWNEEE 0.090 139 DSLIVKGFNV 0.075 405 ISTFHVLIYG 0.075 366 MSLGLLSLLA 0.075 11 KSLSETCLPN 0.075 134 ASLFPDSLIV 0.075 43 KSLTIRLIRC 0.075 303 KQLGLLSFFF 0.075 361 ISFGIMSLGL 0.075 304 QLGLLSFFFA 0.050 107 LLVGKILIDV 0.050 60 SRNPKFASEF 0.050 269 LVYLAGLLAA 0.050 434 LALVLPSIVI 0.050 397 TLGYVALLIS 0.050 364 GIMSLGLLSL 0.050 401 VALLISTFHV 0.050 147 NVVSAWALQL 0.050 189 PIDLGSLSSA 0.050 264 ITLLSLVYLA 0.050 307 LLSFFFAMVH 0.050 310 FFFAMVHVAY 0.050 209 FTLWRGPVVV 0.050 194 SLSSAREIEN 0.050 240 ARNQQSDFYK 0.050 298 WLQCRKQLGL 0.050 440 SIVILDLLQL 0.050 221 SLATFFFLYS 0.050 436 LVLPSIVILD 0.050 406 STFHVLIYGW 0.050

TABLE IX-V2 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 32 RCPPPCPADF 2.000 23 LSLPSSWDYR 1.500 35 PPCPADFFLY 0.625 22 CLSLPSSWDY 0.500 33 CPPPCPADFF 0.250 11 LSLSSGFTPF 0.150 8 ALSLSLSSGF 0.100 13 LSSGFTPFSC 0.075 2 GSPGLQALSL 0.075 28 SWDYRCPPPC 0.050 1 SGSPGLQALS 0.050 36 PCPADFFLYF 0.050 16 GFTPFSCLSL 0.025 12 SLSSGFTPFS 0.020 24 SLPSSWDYRC 0.020 20 FSCLSLPSSW 0.015 14 SSGFTPFSCL 0.015 9 LSLSLSSGFT 0.015 18 TPFSCLSLPS 0.013 7 QALSLSLSSG 0.010 5 GLQALSLSLS 0.010 6 LQALSLSLSS 0.007 10 SLSLSSGFTP 0.005 15 SGFTPFSCLS 0.003 3 SPGLQALSLS 0.003 17 FTPFSCLSLP 0.003 34 PPPCPADFFL 0.001 4 PGLQALSLSL 0.001 31 YRCPPPCPAD 0.001 21 SCLSLPSSWD 0.001 27 SSWDYRCPPP 0.000 25 LPSSWDYRCP 0.000 26 PSSWDYRCPP 0.000 19 PFSCLSLPSS 0.000 30 DYRCPPPCPA 0.000 29 WDYRCPPPCP 0.000

TABLE IX-V5A HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 1 ENLPLRLFTF 1.250 8 FTFWRGPVVV 0.050 3 LPLRLFTFWR 0.013 2 NLPLRLFTFW 0.010 6 RLFTFWRGPV 0.010 7 LFTFWRGPVV 0.001 4 PLRLFTFWRG 0.000 10 FWRGPVVVAI 0.000 5 LRLFTFWRGP 0.000 9 TFWRGPVVVA 0.000

TABLE IX-V5B HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 18 QTELELEFVF 112.500 20 ELELEFVFLL 4.500 22 ELEFVFLLTL 4.500 14 FADTQTELEL 2.500 16 DTQTELELEF 1.250 2 WREFSFIQIF 0.450 5 FSFIQIFCSF 0.150 12 CSFADTQTEL 0.015 9 QIFCSFADTQ 0.010 7 FIQIFCSFAD 0.005 8 IQIFCSFADT 0.003 21 LELEFVFLLT 0.003 4 EFSFIQIFCS 0.003 24 EFVFLLTLLL 0.003 3 REGSFIQIFC 0.003 17 TQTELELEFV 0.002 11 FCSFADTQTE 0.001 19 TELELEFVFL 0.001 6 SFIQIFCSFA 0.001 10 IFCSFADTQT 0.001 23 LEFVFLLTLL 0.001 1 NWREFSFIQI 0.000 15 ADTQTELELE 0.000 13 SFADTQTELE 0.000

TABLE IX-V6 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 42 GTIPHVSPER 5.000 2 VLPSIVILGK 1.000 35 FLEEGIGGTI 0.900 1 LVLPSIVILG 0.500 12 IILFLPCISR 0.500 6 IVILGKIILF 0.500 13 ILFLPCISRK 0.200 15 FLPCISRKLK 0.200 16 LPCISRKLKR 0.125 46 HVSPERVTVM 0.100 7 VILGKIILFL 0.050 5 SIVILGLIIL 0.050 18 CISRKLKRIK 0.020 19 ISRKLKRIKK 0.015 32 KSQFLEEGIG 0.015 39 GIGGTIPHVS 0.010 43 TIPHVSPERV 0.010 11 KIILFLPCIS 0.010 33 SQFLEEGIGG 0.007 38 EGIGGTIPHV 0.005 14 LFLPCISRKL 0.005 36 LEEGIGGTIP 0.005 37 EEGIGGTIPH 0.003 3 LPSIVILGKI 0.003 44 IPHVSPERVT 0.003 29 GWEKSQFLEE 0.002 4 PSIVILGKII 0.002 9 LGKIILFLPC 0.001 23 LKRIKKGWEK 0.001 17 PCISRKLKRI 0.001 10 GKIILFLPCI 0.001 26 IKKGWEKSQF 0.001 34 QFLEEGIGGT 0.001 31 EKSQFLEEGI 0.001 27 KKGWEKSQFL 0.001 8 ILGKIILFLP 0.001 40 IGGTIPHVSP 0.001 41 GGTIPHVSPE 0.000 28 KGWEKSQFLE 0.000 25 RIKKGWEKSQ 0.000 45 PHVSPERVTV 0.000 21 RKLKRIKKGW 0.000 20 SRKLKRIKKG 0.000 30 WEKSQFLEEG 0.000 24 KRIKKGWEKS 0.000 22 KLKRIKKGWE 0.000

TABLE IX-V7A HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 6 LSETFLPNGI 1.350 10 FLPNGINGIK 0.200 8 ETFLPNGING 0.125 4 KSLSETFLPN 0.075 5 SLSETFLPNG 0.020 1 GSPKSLSETF 0.015 9 TFLPNGINGI 0.005 7 SETFLPNGIN 0.001 2 SPKSLSETFL 0.000 3 PKSLSETFLP 0.000

TABLE IX-V7B HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 MAYQQSTLGY 2.500 10 STLGYVALLI 0.125 9 QSTLGYVALL 0.030 2 FLNMAYQQST 0.010 4 NMAYQQSTLG 0.005 7 YQQSTLGYVA 0.003 8 QQSTLGYVAL 0.003 3 LNMAYQQSTL 0.003 6 AYQQSTLGYV 0.001 1 LFLNMAYQQS 0.001

TABLE IX-V7C HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 100 SIDPPESPDR 100.000 67 TAEAQESGIR 9.000 33 LSEIVLPIEW 6.750 131 LWEFLLRLLK 4.500 91 VTEDDEAQDS 2.250 10 SVEVLASPAA 1.800 52 STPPPPAMWT 1.250 6 ILDLSVEVLA 1.000 168 KLETIILSKL 0.900 103 PPESPDRALK 0.900 127 GVGPLWEFLL 0.500 143 AASGTLSLAF 0.500 13 VLASPAAAWK 0.400 51 LSTPPPPAMW 0.300 60 WTEEAGATAE 0.225 157 LGEFLGSGTW 0.225 69 EAQESGIRNK 0.200 97 AQDSIDPPES 0.150 70 AQESGIRNKS 0.135 178 TQEQKSKHCM 0.135 170 ETIILSKLTQ 0.125 128 VGPLWEFLLR 0.125 37 VLPIEWQQDR 0.100 14 LASPAAAWKC 0.100 61 TEEAGATAEA 0.090 39 PIEWQQDRKI 0.090 162 GSGTWMKLET 0.075 78 KSSSSSQIPV 0.075 160 FLGSGTWMKL 0.050 22 KCLGANILRG 0.050 167 MKLETIILSK 0.050 38 LPIEWQQDRK 0.050 80 SSSSQIPVVG 0.030 79 SSSSSQIPVV 0.030 83 SQIPVVGVVT 0.030 144 ASGTLSLAFT 0.030 81 SSSQIPVVGV 0.030 146 GTLSLAFTSW 0.025 66 ATAEAQESGI 0.025 152 FTSWSLGEFL 0.025 125 TNGVGPLWEF 0.025 92 TEDDEAQDSI 0.025 177 LTQEQKSKHC 0.025 21 WKCLGANILR 0.025 106 SPDRALKAAN 0.025 94 DDEAQDSIDP 0.022 12 EVLASPAAAW 0.020 4 IVILDLSVEV 0.020 173 ILSKLTQEQK 0.020 47 KIPPLSTPPP 0.020 113 AANSWRNPVL 0.020 72 ESGIRNKSSS 0.015 43 QQDRKIPPLS 0.015 15 ASPAAAWKCL 0.015 140 KSQAASGTLS 0.015 9 LSVEVLASPA 0.015 82 SSQIPVVGVV 0.015 155 WSLGEFLGSG 0.015 105 ESPDRALKAA 0.015 148 LSLAFTSWSL 0.015 124 HTNGVGPLWE 0.013 129 GPLWEFLLRL 0.013 31 GGLSEIVLPI 0.013 145 SGTLSLAFTS 0.013 185 HCMFSLISGS 0.010 149 SLAFTSWSLG 0.010 65 GATAEAQESG 0.010 112 KAANSWRNPV 0.010 142 QAASGTLSLA 0.010 25 GANILRGGLS 0.010 159 EFLGSGTWMK 0.010 23 CLGANILRGG 0.010 109 RALKAANSWR 0.010 176 KLTQEQKSKH 0.010 35 EIVLPIEWQQ 0.010 175 SKLTQEQKSK 0.010 18 AAAWKCLGAN 0.010 36 IVLPIEWQQD 0.010 5 VILDLSVEVL 0.010 172 IILSKLTQEQ 0.010 156 SLGEFLGSGT 0.010 120 PVLPHTNGVG 0.010 147 TLSLAFTSWS 0.010 89 GVVTEDDEAQ 0.010 153 TSWSLGEFLG 0.008 2 PSIVILDLSV 0.008 141 SQAASGTLSL 0.007 150 LAFTSWSLGE 0.005 17 PAAAWKCLGA 0.005 101 IDPPESPDRA 0.005 151 AFTSWSLGEF 0.005 117 WRNPVLPHTN 0.005 42 WQQDRKIPPL 0.003 104 PESPDRALKA 0.003 24 LGANILRGGL 0.003 119 NPVLPHTNGV 0.003 118 RNPVLPHTNG 0.003 102 DPPESPDRAL 0.003 53 TPPPPAMWTE 0.003 1 LPSIVILDLS 0.003

TABLE X-V1 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 227 FLYSFVRDV 1789.612 402 ALLISTFHV 1492.586 307 LLSFFFAMV 853.681 306 GLLSFFFAM 769.748 100 SLWDLRHLL 726.962 333 FLNMAYQQV 479.909 140 SLIVKGFNV 403.402 203 NLPLRLFTL 284.974 210 TLWRGPVVV 236.685 65 FASEFFPHV 131.539 135 SLFPDSLIV 105.510 274 GLLAAAYQL 79.041 393 FIQSTLGYV 72.344 48 RLIRCGYHV 69.552 365 IMSLGLLSL 60.325 5 SMMGSPKSL 57.085 220 ISLATFFFL 53.163 271 YLAGLLAAA 52.561 265 TLLSLVYLA 42.278 433 VLALVLPSI 40.792 442 VILDLLQLC 40.518 112 ILIDVSNNM 34.627 360 YISFGIMSL 31.077 403 LLISTFHVL 28.290 369 GLLSLLAVT 26.001 17 CLPNGINGI 23.995 108 LVGKILIDV 23.795 264 ITLLSLVYL 23.608 258 TLPIVAITL 21.362 184 QLNFIPIDL 21.362 313 AMVHVAYSL 15.428 410 VLIYGWKRA 14.358 141 LIVKGFNVV 12.665 305 LGLLSFFFA 12.364 44 SLTIRLIRC 11.426 436 LVLPSIVIL 11.087 397 TLGYVALLI 10.433 386 LNWREFSFI 10.042 180 ELARQLNFI 9.898 254 IVNKTLPIV 9.756 404 LISTFHVLI 9.267 357 IEMYISFGI 7.401 441 IVILDLLQL 7.309 261 IVAITLLSL 7.309 209 FTLWRGPVV 6.741 368 LGLLSLLAV 6.568 367 SLGLLSLLA 4.968 153 ALQLGPKDA 4.968 146 FNVVSAWAL 4.811 389 REFSFIQST 4.686 435 ALVLPSIVI 4.277 187 FIPIDLGSL 4.040 374 LAVTSIPSV 3.777 262 VAITLLSLV 3.777 299 LQCRKQLGL 3.682 335 NMAYQQVHA 3.588 291 FPPWLETWL 3.528 331 YLFLNMAYQ 3.209 148 VVSAWALQL 3.178 166 YICSNNIQA 3.142 353 EVWRIEMYI 3.125 221 SLATFFFLY 3.121 378 SIPSVSNAL 2.937 164 QVYICSNNI 2.921 268 SLVYLAGLL 2.777 396 STLGYVALL 2.525 434 LALVLPSIV 2.491 304 QLGLLSFFF 2.377 269 LVYLAGLLA 2.365 37 GSGDFAKSL 2.173 366 MSLGLLSLL 2.017 267 LSLVYLAGL 2.017 242 NQQSDFYKI 2.010 177 QVIELARQL 1.533 224 TFFFLYSFV 1.474 349 WNEEEVWRI 1.418 128 SNAEYLASL 1.315 106 HLLVGKILI 1.312 257 KTLPIVAIT 1.264 303 KQLGLLSFF 1.238 428 TPPNFVLAL 1.219 34 GVIGSGDFA 1.172 216 VVVAISLAT 1.108 314 MVHVAYSLC 1.108 371 LSLLAVTSI 0.985 91 VAIHREHYT 0.968 85 KTNIIFVAI 0.964 133 LASLFPDSL 0.939 425 RFYTPPNFV 0.850 250 IPIEIVNKT 0.780 49 KIRCGYHVV 0.760 83 LTKTNIIFV 0.727 132 YLASLFPDS 0.651 427 YTPPNFVLA 0.603 171 NIQARQQVI 0.588 259 LPIVAITLL 0.545 438 LPSIVILDL 0.545 278 AAYQLYYGT 0.497 170 NNIQARQQV 0.454 385 ALNWREFSF 0.432

TABLE X-V2 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 5 GLQALSLSL 21.362 10 SLSLSSGFT 5.328 17 FTPFSCLSL 1.365 15 SGFTPFSCL 0.980 1 SGSPGLQAL 0.321 14 SSGFTPFSC 0.188 8 ALSLSLSSG 0.171 12 SLSSGFTPF 0.142 3 SPGLQALSL 0.139 29 WDYRCPPPC 0.102 35 PPCPADFFL 0.098 22 CLSLPSSWD 0.082 37 CPADFFLYF 0.079 24 SLPSSWDYR 0.068 25 LPSSWDYRC 0.055 6 LQALSLSLS 0.030 23 LSLPSSWDY 0.023 13 LSSGFTPFS 0.017 20 FSCLSLPSS 0.005 7 QALSLSLSS 0.004 11 LSLSSGFTP 0.004 27 SSWDYRCPP 0.003 31 YRCPPPCPA 0.003 9 LSLSLSSGF 0.003 21 SCLSLPSSW 0.002 18 TPFSCLSLP 0.001 2 GSPGLQALS 0.000 33 CPPPCPADF 0.000 16 GFTPFSCLS 0.000 36 PCPADFFLY 0.000 32 RCPPPCPAD 0.000 4 PGLQALSLS 0.000 34 PPPCPADFF 0.000 19 PFSCLSLPS 0.000 28 SWDYRCPPP 0.000 26 PSSWDYRCP 0.000 30 DYRCPPPCP 0.000

TABLE X-V5A HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 7 FTFWRGPVV 6.741 1 NLPLRLFTF 0.994 8 TFWRGPVVV 0.164 5 RLFTFWRGP 0.071 2 LPLRLFTFW 0.032 6 LFTFWRGPV 0.011 3 PLRLFTFWR 0.003 4 LRLFTFWRG 0.001 9 FWRGPVVVA 0.000

TABLE X-V5B HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 20 LELEFVFLL 543.025 6 FIQIFCSFA 65.673 24 FVFLLTLLL 31.814 22 LEFVFLLTL 22.835 8 QIFCSFADT 7.203 19 ELELEFVFL 1.072 17 QTELELEFV 0.383 10 FCSFADTQT 0.224 4 FSFIQIFCS 0.110 21 ELEFVFLLT 0.068 12 SFADTQTEL 0.061 18 TELELEFVF 0.052 16 TQTELELEF 0.031 14 ADTQTELEF 0.030 2 REFSFIQIF 0.019 7 IQIFCSFAD 0.015 23 EFVFLLTLL 0.003 3 EFSFIQIFC 0.001 1 WREFSFIQI 0.001 11 CSFADTQTE 0.000 13 FADTQTELE 0.000 5 SFIQIFCSF 0.000 9 IFCSFADTQ 0.000 15 DTQTELELE 0.000

TABLE X-V6 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 7 ILGKIILFL 459.398 27 KGWEKSQFL 91.350 10 KIILFLPCI 43.882 38 GIGGTIPHV 21.996 14 FLPCISRKL 19.653 17 CISRKLKRI 3.299 34 FLEEGIGGT 2.689 5 IVILGKIIL 1.303 4 SIVILGKII 0.588 43 IPHVSPERV 0.378 1 VLPSIVILG 0.291 46 VSPERVTVM 0.213 45 HVSPERVTV 0.207 6 VILGKIILF 0.148 31 KSQFLEEGI 0.117 12 ILFLPCISR 0.094 11 IILFLPCIS 0.026 9 GKIILFLPC 0.013 21 KLKRIKKGW 0.009 35 LEEGIGGTI 0.003 42 TIPHVSPER 0.002 32 SQFLEEGIG 0.001 20 RKLKRIKKG 0.001 33 QFLEEGIGG 0.001 41 GTIPHVSPE 0.000 3 PSIVILGKI 0.000 2 LPSIVILGK 0.000 26 KKGWEKSQF 0.000 39 IGGTIPHVS 0.000 24 RIKKGWEKS 0.000 15 LPCISRKLK 0.000 13 LFLPCISRK 0.000 40 GGTIPHVSP 0.000 29 WEKSQFLEE 0.000 8 LGKIILFLP 0.000 23 KRIKKGWEK 0.000 37 EGIGGTIPH 0.000 30 EKSQFLEEG 0.000 44 PHVSPERVT 0.000 36 EEGIGGTIP 0.000 16 PCISRKLKR 0.000 22 LKRIKKGWE 0.000 25 IKKGWEKSQ 0.000 18 ISRKLKRIK 0.000 28 GWEKSQFLE 0.000 19 SRKLKRIKK 0.000

TABLE X-V7A HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 FLPNGINGI 110.379 4 SLSETFLPN 0.581 6 SETFLPNGI 0.203 3 KSLSETFLP 0.007 2 PKSLSETFL 0.004 5 LSETFLPNG 0.000 8 TFLPNGING 0.000 7 ETFLPNGIN 0.000 1 SPKSLSETF 0.000

TABLE X-V7B HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 6 YQQSTLGYV 53.345 3 NMAYQQSTL 15.428 9 STLGYVALL 2.525 1 FLNMAYQQS 0.514 2 LNMAYQQST 0.306 8 QSTLGYVAL 0.209 7 QQSTLGYVA 0.207 4 MAYQQSTLG 0.006 5 AYQQSTLGY 0.000

TABLE X-V7C A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 4 VILDLSVEV 246.631 148 SLAFTSWSL 160.218 129 PLWEFLLRL 139.780 31 GLSEIVLPI 98.381 57 AMWTEEAGA 29.780 2 SIVILDLSV 9.563 126 GVGPLWEFL 8.564 5 ILDLSVEVL 6.712 152 TSWSLGEFL 3.119 27 ILRGGLSEI 3.100 42 QQDRKIPPL 1.993 168 LETIILSKL 1.624 127 VGPLWEFLL 1.375 163 GTWMKLETI 1.355 81 SSQIPVVGV 1.044 165 WMKLETIIL 1.018 112 AANSWRNPV 0.966 82 SQIPVVGVV 0.864 134 LLRLLKSQA 0.642 144 SGTLSLAFT 0.615 133 FLLRLLKSQ 0.583 39 IEWQQDRKI 0.572 159 FLGSGTWMK 0.514 119 PVLPHTNGV 0.495 185 CMFSLISGS 0.458 78 SSSSSQIPV 0.454 79 SSSSQIPVV 0.428 83 QIPVVGVVT 0.420 160 LGSGTWMKL 0.403 155 SLGEFLGSG 0.347 141 QAASGTLSL 0.297 136 RLLKSQAAS 0.276 52 TPPPPAMWT 0.268 14 ASPAAAWKC 0.243 15 SPAAAWKCL 0.237 181 KSKHCMFSL 0.228 88 GVVTEDDEA 0.213 22 CLGANILRG 0.171 10 VEVLASPAA 0.164 142 AASGTLSLA 0.159 146 TLSLAFTSW 0.142 12 VLASPAAAW 0.127 11 EVLASPAAA 0.121 49 PLSTPPPPA 0.109 178 QEQKSKHCM 0.097 59 WTEEAGATA 0.083 17 AAAWKCLGA 0.069 147 LSLAFTSWS 0.064 139 KSQAASGTL 0.063 35 IVLPIEWQQ 0.062 29 RGGLSEIVL 0.057 113 ANSWRNPVL 0.057 20 WKCLGANIL 0.056 50 LSTPPPPAM 0.055 175 KLTQEQKSK 0.052 162 SGTWMKLET 0.049 6 LDLSVEVLA 0.043 36 VLPIEWQQD 0.043 24 GANILRGGL 0.039 177 TQEQKSKHC 0.032 105 SPDRALKAA 0.030 171 IILSKLTQE 0.030 41 WQQDRKIPP 0.028 9 SVEVLASPA 0.028 182 SKHCMFSLI 0.028 172 ILSKLTQEQ 0.025 145 GTLSLAFTS 0.022 138 LKSQPASGT 0.018 154 WSLGEFLGS 0.016 76 NKSSSSSQI 0.014 7 DLSVEVLAS 0.013 149 LAFTSWSLG 0.011 116 WRNPVLPHT 0.011 104 ESPDRALKA 0.010 66 TAEAQESGI 0.009 125 NGVGPLWEF 0.008 169 ETIILSKLT 0.008 167 KLETIILSK 0.008 26 NILRGGLSE 0.008 140 SQAASGTLS 0.008 61 EEAGATAEA 0.007 176 LTQEQKSKH 0.007 46 KIPPLSTPP 0.007 120 VLPHTNGVG 0.007 166 MKLETIILS 0.006 156 LGEFLGSGT 0.005 158 EFLGSGTWM 0.005 131 WEFLLRLLK 0.005 101 DPPESPDRA 0.005 89 VVTEDDEAQ 0.004 137 LLKSQAASG 0.004 135 LRLLKSQAA 0.004 108 RALKAANSW 0.004 28 LRGGLSEIV 0.003 109 ALKAANSWR 0.003 18 AAWKCLGAN 0.003 91 TEDDEAQDS 0.002 164 TWMKLETII 0.002 3 IVILDLSVE 0.002 65 ATAEAQESG 0.002

TABLE XI-V1 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 100 SLWDLRHLLV 2366.855 306 GLLSFFFAMV 1858.012 82 ALTKTNIIFV 879.833 304 QLGLLSFFFA 301.110 373 LLAVTSIPSV 271.948 107 LLVGKILIDV 271.948 132 YLASLFPDSL 182.973 219 AISLATFFFL 178.032 367 SLGLLSLLAV 159.970 385 ALNWREFSFI 109.023 298 WLQCRKQLGL 98.267 437 VLPSIVILDL 83.527 266 LLSLVYLAGL 83.527 403 LLISTFHVLI 67.396 402 ALLISTFHVL 61.573 365 IMSLGLLSLL 60.325 140 SLIVKGFNVV 54.181 258 TLPIVAITLL 49.134 433 VLALVLPSIV 48.478 48 RLIRCGYHVV 42.774 370 LLSLLAVTSI 40.792 210 TLWRGPVVVA 38.884 263 AITLLSLVYL 37.157 432 FVLALVLPSI 35.735 401 VALLISTFHV 35.242 207 RLFTLWRGPV 33.455 277 FLYSFVRDVI 30.852 223 ATFFFLYSFV 29.487 65 FASEFFPHVV 28.385 364 GIMSLGLLSL 24.997 261 IVAITLLSLV 23.795 435 ALVLPSIVIL 20.145 90 FVAIHREHYT 16.497 179 IELARQLNFI 16.141 427 YTPPVFVLAL 11.929 67 SEFFPHVVDV 11.509 111 KILIDVSNNM 8.846 305 LGLLSFFFAM 8.542 172 IQARQQVIEL 8.469 249 KIPIEIVNKT 8.248 183 RQLNFIPIDL 8.014 95 REHYTSLWDL 7.165 440 SIVILDLLQL 6.756 209 FTLWRGPVVV 6.741 308 LSFFFAMVHV 6.568 57 VIGSRNPKFA 6.387 419 FEEEEYRFYT 5.579 394 IQSTLGYVAL 5.523 269 LVYLAGLLAA 5.439 313 AMVHVAYSLC 5.382 312 FAMVHVAYSL 5.050 268 SLVYLAGLLA 4.968 92 AIHREHYTSL 4.406 243 QQSDFYKIPI 4.337 257 KTLPIVAITL 3.842 231 FVRDVIHPYA 3.427 314 MVHVAYSLCL 3.178 303 KQLGLLSFFF 3.121 221 SLATFFFLYS 2.959 144 KGFNVVSAWA 2.310 286 TKYRRFPPWL 1.984 147 NVVSAWALQL 1.869 199 REIENLPLRL 1.703 441 IVILDLLQLC 1.700 389 REFSFIQSTL 1.537 226 FFLYSFVRDV 1.437 24 GIKDARKVTV 1.372 201 IENLPLRLFT 1.355 393 FIQSTLGYVA 1.288 64 KFASEFFPHV 1.221 152 WALQLGPKDA 1.174 345 IENSWNEEEV 1.127 299 LQCRKQLGLL 1.101 163 RQVYICSNNI 1.058 428 TPPNFVLALV 1.044 264 ITLLSLVYLA 0.998 113 LIDVSNNMRI 0.975 250 IPIEIVNKTL 0.972 43 KSLTIRLIRC 0.966 323 LPMRRSERYL 0.965 424 YRFYTPPNFV 0.904 36 IGSGDFAKSL 0.901 361 ISFGIMSLGL 0.877 4 ISMMGSPKSL 0.877 336 MAYQQVHANI 0.788 139 DSLIVKGFNV 0.731 12 SLSETCLPNG 0.703 275 LLAAAYQLYY 0.697 134 ASLFPDSLIV 0.689 121 RINQYPESNA 0.683 253 EIVNKTLPIV 0.676 98 YTSLWDLRHL 0.628 398 LGYVALLIST 0.609 16 TCLPNGINGI 0.580 396 STLGYVALLI 0.536 356 RIEMYISFGI 0.532 202 ENLPLRLFTL 0.516 99 TSLWDLRHLL 0.516 273 AGLLAAAYQL 0.516 332 LFLNMAYQQV 0.456

TABLE XI-V2 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 24 SLPSSWDYRC 4.968 12 SLSSGFTPFS 1.557 22 CLSLPSSWDY 0.559 13 LSSGFTPFSC 0.320 14 SSGFTPFSCL 0.265 9 LSLSLSSGFT 0.219 5 GLQALSLSLS 0.171 2 GSPGLQALSL 0.139 34 PPPCPADFFL 0.098 10 SLSLSSGFTP 0.086 8 ALSLSLSSGF 0.075 16 GFTPFSCLSL 0.015 6 LQALSLSLSS 0.013 4 PGLQALSLSL 0.011 7 QALSLSLSSG 0.009 15 SGFTPFSCLS 0.007 11 LSLSSGFTPF 0.006 27 SSWDYRCPPP 0.003 23 LSLPSSWDYR 0.003 20 FSCLSLPSSW 0.002 17 FTPFSCLSLP 0.002 21 SCLSLPSSWD 0.002 18 TPFSCLSLPS 0.002 33 CPPPCPADFF 0.001 3 SPGLQALSLS 0.001 32 RCPPPCPADF 0.000 1 SGSPGLQALS 0.000 36 PCPADFFLYF 0.000 29 WDYRCPPPCP 0.000 28 SWDYRCPPPC 0.000 35 PPCPADFFLY 0.000 25 LPSSWDYRCP 0.000 31 YRCPPPCPAD 0.000 30 DYRCPPPCPA 0.000 19 PFSCLSLPSS 0.000 26 PSSWDYRCPP 0.000

TABLE XI-V5A HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 6 RLFTFWRGPV 33.455 8 FTFWRGPVVV 6.741 2 NLPLRLFTFW 0.779 3 LPLRLFTFWR 0.074 7 LFTFWRGPVV 0.034 9 TFWRGPVVVA 0.027 1 ENLPLRLFTF 0.002 4 PLRLFTFWRG 0.002 10 FWRGPVVVAI 0.001 5 LRLFTFWRGP 0.000

TABLE XI-V5B HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 17 TQTELELEFV 179.213 19 TELELEFVFL 65.849 21 LELEFVFLLT 7.100 23 LEFVFLLTLL 6.009 20 ELELEFVFLL 5.198 8 IQIFCSFADT 2.440 3 REFSFIQIFC 1.966 22 ELEFVFLLTL 0.896 14 FADTQTELEL 0.546 12 CSFADTQTEL 0.516 6 SFIQIFCSFA 0.072 7 FIQIFCSFAD 0.055 5 FSFIQIFCSF 0.016 9 QIFCSFADTQ 0.014 10 IFCSFADTQT 0.009 24 EFVFLLTLLL 0.001 1 NWREFSFIQI 0.001 11 FCSFADTQTE 0.000 18 QTELELEFVF 0.000 16 DTQTELELEF 0.000 4 EFSFIQIFCS 0.000 15 ADTQTELELE 0.000 13 SFADTQTELE 0.000 2 WREFSFIQIF 0.000

TABLE XI-V6 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 7 VILGKIILFL 233.719 43 TIPHVSPERV 4.686 35 FLEEGIGGTI 1.637 5 SIVILGKIIL 1.204 27 KKGWEKSQFL 0.571 8 ILGKIILFLP 0.338 13 ILFLPCISRK 0.216 10 GKIILFLPCI 0.127 1 LVLPSIVILG 0.094 38 EGIGGTIPHV 0.078 15 FLPCISRKLK 0.069 28 KGWEKSQFLE 0.067 2 VLPSIVILGK 0.058 3 LPSIVILGKI 0.035 33 SQFLEEGIGG 0.028 6 IVILGKIILF 0.025 34 QFLEEGIGGT 0.023 14 LFLPCISRKL 0.019 11 KIILFLPCIS 0.015 46 HVSPERVTVM 0.014 12 IILFLPCISR 0.013 44 IPHVSPERVT 0.007 39 GIGGTIPHVS 0.004 9 LGKIILFLPC 0.004 17 PCISRKLKRI 0.003 22 KLKRIKKGWE 0.001 45 PHVSPERVTV 0.001 30 WEKSQFLEEG 0.001 4 PSIVILGKII 0.001 31 EKSQFLEEGI 0.001 21 RKLKRIKKGW 0.000 41 GGTIPHVSPE 0.000 42 GTIPHVSPER 0.000 18 CISRKLKRIK 0.000 40 IGGTIPHVSP 0.000 16 LPCISRKLKR 0.000 37 EEGIGGTIPH 0.000 32 KSQFLEEGIG 0.000 25 RIKKGWEKSQ 0.000 24 KRIKKGWEKS 0.000 23 LKRIKKGWEK 0.000 36 LEEGIGGTIP 0.000 19 ISRLKLRIKK 0.000 26 IKKGWEKSQF 0.000 20 SRKLKRIKKG 0.000 29 GWEKSQFLEE 0.000

TABLE XI-V7A HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 SLSETFLPNG 2.670 9 TFLPNGINGI 0.062 2 SPKSLSETFL 0.027 4 KSLSETFLPN 0.012 6 LSETFLPNGI 0.007 10 FLPNGINGIK 0.004 8 ETFLPNGING 0.000 1 GSPKSLSETF 0.000 7 SETFLPNGIN 0.000 3 PKSLSETFLP 0.000

TABLE XI-V7B HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 2 FLNMAYQQST 34.279 8 QQSTLGYVAL 3.249 7 YQQSTLGYVA 0.950 3 LNMAYQQSTL 0.877 10 STLGYVALLI 0.536 9 QSTLGYVALL 0.321 4 NMAYQQSTLG 0.054 6 AYQQSTLGYV 0.016 5 MAYQQSTLGY 0.006 1 LFLNMAYQQS 0.000

TABLE XI-V7C HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 160 FLGSGTWMKL 167.054 42 WQQDRKIPPL 93.953 134 FLLRLLKSQA 84.555 5 VILDLSVEVL 35.002 156 SLGEFLGSGT 30.553 27 NILRGGLSEI 12.208 168 KLETIILSKL 11.006 127 GVGPLWEFLL 10.841 4 IVILDLSVEV 10.346 130 PLWEFLLRLL 7.357 148 LSLAFTSWSL 6.579 58 AMWTEEAGAT 5.807 129 GPLWEFLLRL 4.510 152 FTSWSLGEFL 3.678 112 KAANSWRNPV 3.381 6 ILDLSVEVLA 3.378 141 SQAASGTLSL 2.166 158 GEFLGSGTWM 1.966 28 ILRGGLSEIV 1.805 78 KSSSSSQIPV 1.589 147 TLSLAFTSWS 1.557 19 AAWKCLGANI 1.203 81 SSSQIPVVGV 1.044 14 LASPAAAWKC 0.880 135 LLRLLKSQAA 0.642 126 NGVGPLWEFL 0.639 144 ASGTLSLAFT 0.615 66 ATAEAQESGI 0.594 31 GGLSEIVLPI 0.580 52 STPPPPAMWT 0.569 164 GTWMKLETII 0.493 177 LTQEQKSKHC 0.481 119 NPVLPHTNGV 0.454 138 LLKSQAASGT 0.443 79 SSSSSQIPVV 0.428 181 QKSKHCMFSL 0.396 83 SQIPVVGVVT 0.310 137 RLLKSQAASG 0.276 176 KLTQEQKSKH 0.261 169 LETIILSKLT 0.246 15 ASPAAAWKCL 0.237 9 LSVEVLASPA 0.226 11 VEVLASPAAA 0.164 92 TEDDEAQDSI 0.163 142 QAASGTLSLA 0.159 13 VLASPAAAWK 0.139 149 SLAFTSWSLG 0.127 113 AANSWRNPVL 0.122 50 PLSTPPPPAM 0.109 163 SGTWMKLETI 0.077 122 LPHTNGVGPL 0.071 32 GLSEIVLPIE 0.058 132 WEFLLRLLKS 0.057 82 SSQIPVVGVV 0.056 162 GSGTWMKLET 0.049 23 CLGANILRGG 0.034 178 TQEQKSKHCM 0.032 24 LGANILRGGL 0.031 10 SVEVLASPAA 0.028 88 VGVVTEDDEA 0.027 37 VLPIEWQQDR 0.025 121 VLPHTNGVGP 0.025 153 TSWSLGEFLG 0.023 105 ESPDRALKAA 0.023 166 WMKLETIILS 0.020 110 ALKAANSWRN 0.020 182 KSKHCMFSLI 0.016 22 KCLGANILRG 0.014 36 IVLPIEWQQD 0.014 172 IILSKLTQEQ 0.013 173 ILSKLTQEQK 0.012 2 PSIVILDLSV 0.010 155 WSLGEFLGSG 0.009 115 NSWRNPVLPH 0.009 90 VVTEDDEAQD 0.009 102 DPPESPDRAL 0.009 125 TNGVGPLWEF 0.008 146 GTLSLAFTSW 0.007 47 KIPPLSTPPP 0.007 139 LKSQAASGTL 0.007 61 TEEAGATAEA 0.006 101 IDPPESPDRA 0.006 57 PAMWTEEAGA 0.006 59 MWTEEAGATA 0.005 171 TIILSKLTQE 0.005 84 QIPVVGVVTE 0.005 165 TWMKLETIIL 0.005 109 RALKAANSWR 0.004 97 AQDSIDPPES 0.003 43 QQDRKIPPLS 0.003 145 SGTLSLAFTS 0.003 49 PPLSTPPPPA 0.003 8 DLSVEVLASP 0.003 76 RNKSSSSSQI 0.002 104 PESPDRALKA 0.002 29 LRGGLSEIVL 0.002 3 SIVILDLSVE 0.002 12 EVLASPAAAW 0.002 34 SEIVLPIEWQ 0.002 140 KSQAASGTLS 0.002

TABLE XII-V1 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 221 SLATEFFLY 108.000 306 GLLSFFFAM 24.300 294 WLETWLQCR 18.000 281 QLYYGTKYR 10.000 249 KIPIEIVNK 9.000 103 DLRHLLVGK 9.000 274 GLLAAAYQL 8.100 443 ILDLLQLCR 8.000 223 ATFFFLYSF 6.750 304 QLGLLSFFF 6.000 155 QLGPKDASR 6.000 385 ALNWREFSF 6.000 35 VIGSGDFAK 6.000 409 HVLIYGWKR 5.400 56 IWIGSRNPK 4.500 313 AMVHVAYSL 4.050 82 ALTKTNIIF 4.000 322 CLPMRRSER 4.000 275 LLAAAYQLY 4.000 135 SLFPDSLIV 3.000 100 SLWDLRHLL 3.000 21 GINGIKDAR 2.700 403 LLISTFHVL 2.700 265 TLLSLVYLA 2.700 435 ALVLPSIVI 2.700 203 NLPLRLFTL 2.700 205 PLRLFTLWR 2.400 3 SISMMGSPK 2.000 258 TLPIVAITL 1.800 184 QLNFIPIDL 1.800 397 TLGYVALLI 1.800 365 IMSLGLLSL 1.800 307 LLSFFFAMV 1.800 87 NIIFVAIHR 1.800 106 HLLVGKILI 1.800 433 VLALVLPSI 1.350 191 DLGSLSSAR 1.200 210 TLWRGPVVV 1.000 140 SLIVKGFNV 0.900 17 CLPNGINGI 0.900 231 FVRDVIHPY 0.900 48 RLIRCGYHV 0.900 402 ALLISTFHV 0.900 227 FLYSFVRDV 0.900 417 RAFEEEYYR 0.900 263 AITLLSLVY 0.800 5 SMMGSPKSL 0.675 369 GLLSLLAVT 0.675 396 STLGYVALL 0.608 303 KQLGLLSFF 0.608 44 SLTIRLIRC 0.600 381 SVSNALNWR 0.600 46 TIRLIRCGY 0.600 219 AISLATFFF 0.600 280 YQLYYGTKY 0.540 411 LIYGWKRAF 0.450 271 YLAGLLAAA 0.450 112 ILIDVSNNM 0.450 85 KTNIIFVAI 0.405 98 FVAIHREHY 0.400 367 SLGLLSLLA 0.400 113 LIDVSNNMR 0.400 148 VVSAWALQL 0.360 175 RQQVIELAR 0.360 217 VVAISLATF 0.300 164 QVYICSNNI 0.300 400 YVALLISTF 0.300 43 KSLTIRLIR 0.270 441 IVILDLLQL 0.270 268 SLVYLAGLL 0.270 180 ELARQLNFI 0.270 353 EVWRIEMYI 0.270 358 EMYISFGIM 0.270 276 LAAAYQLYY 0.240 436 LVLPSIVIL 0.203 335 NMAYQQVHA 0.200 57 VIGSRNPKF 0.200 269 LVYLAGLLA 0.200 333 FLNMAYQQV 0.200 261 IVAITLLSL 0.180 225 FFFLYSFVR 0.180 360 YISFGIMSL 0.180 437 VLPSIVILD 0.180 404 LISTFHVLI 0.180 242 NQQSDFYKI 0.162 257 KTLPIVAIT 0.152 331 YLFLNMAYQ 0.150 410 VLIYGWKRA 0.150 34 GVIGSGDFA 0.135 18 LPNGINGIK 0.135 107 LLVGKILID 0.135 241 RNQQSDFYK 0.120 405 ISTFHVLIY 0.120 132 YLASLFPDS 0.120 428 TPPNFVLAL 0.108 153 ALQLGPKDA 0.100 108 LVGKILIDV 0.090 378 SIPSVSNAL 0.090 141 LIVKGFNVV 0.090 282 LYYGTKYRR 0.090

TABLE XII-V2 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 12 SLSSGFTPF 6.000 24 SLPSSWDYR 4.000 5 GLQALSLSL 3.600 37 CPADFFLYF 0.360 23 LSLPSSWDY 0.135 17 FTPFSCLSL 0.060 36 PCPADFFLY 0.036 8 ALSLSLSSG 0.030 22 CLSLPSSWD 0.030 10 SLSLSSGFT 0.030 33 CPPPCPADF 0.030 25 LPSSWDYRC 0.018 9 LSLSLSSGF 0.015 15 SGFTPFSCL 0.013 3 SPGLQALSL 0.012 34 PPPCPADFF 0.003 14 SSGFTPFSC 0.003 21 SCLSLPSSW 0.003 35 PPCPADFFL 0.003 6 LQALSLSLS 0.002 18 TPFSCLSLP 0.002 27 SSWDYRCPP 0.002 1 SGSPGLQAL 0.001 7 QALSLSLSS 0.001 29 WDYRCPPPC 0.001 13 LSSGFTPFS 0.001 2 GSPGLQALS 0.001 16 GFTPFSCLS 0.001 31 YRCPPPCPA 0.000 11 LSLSSGFTP 0.000 32 RCPPPCPAD 0.000 20 FSCLSLPSS 0.000 28 SWDYRCPPP 0.000 4 PGLQALSLS 0.000 30 DYRCPPPCP 0.000 19 PFSCLSLPS 0.000 26 PSSWDYRCP 0.000

TABLE XII-V5A HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 NLPLRLFTF 9.000 3 PLRLFTFWR 3.600 7 FTFWRGPVVW 0.050 5 RLFTFWRGP 0.030 2 LPLRLFTFW 0.009 9 FWRGPVVVA 0.001 8 TFWRGPVVV 0.001 4 LRLFTFWRG 0.000 6 LFTFWRGPV 0.000

TABLE XII-V5B HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 24 FVFLLTLLL 0.600 19 ELELEFVFL 0.540 21 ELEFVFLLT 0.270 16 TDTELELEF 0.180 8 QIFCSFADT 0.150 2 REFSFIQIF 0.135 20 LELEFVFLL 0.109 22 LEEVFLLTL 0.081 6 FIQIFCSFA 0.060 18 TELELEEVE 0.041 17 QTELELEFV 0.015 5 SFIQIFCSF 0.013 4 FSFIQIFCS 0.005 1 WREFSFIQI 0.004 7 IQIFCSFAD 0.003 14 ADTQTELEL 0.001 10 FCSFADTQT 0.001 12 SFADTQTEL 0.001 11 CSFADTQTE 0.001 15 DTQTELELE 0.000 23 EFVFLLTLL 0.000 13 FADTQTELE 0.000 3 EFSFIQIFC 0.000 9 IFCSFADTQ 0.000

TABLE XII-V6 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 12 ILFLPCISR 60.000 7 ILGKIILFL 2.700 6 VILGKIILF 1.350 10 KIILELPCI 1.215 2 LPSIVILGK 0.900 42 TIPHVSPER 0.600 21 KLKRIKKGW 0.450 23 KRIKKGWEK 0.270 5 IVILGKIIL 0.180 1 VLPSIVILG 0.180 38 GIGGTIPHV 0.135 15 LPCISRKLK 0.100 14 FLPCISRKL 0.090 13 LFLPCISRK 0.068 34 FLEEGIGGT 0.068 17 CISRKLKRI 0.045 4 SIVILGKII 0.045 19 SRKLKRIKK 0.040 45 HVSPERVTV 0.030 41 GTIPHVSPE 0.020 27 KGWEKSQFL 0.014 16 PCISRKLKR 0.012 18 ISRKLKRIK 0.010 31 KSQFLEEGI 0.009 26 KKGWEKSQF 0.006 11 IILFLPCIS 0.006 9 GKIILFLPC 0.005 46 VSPERVTVM 0.005 24 RIKKGWEKS 0.004 43 IPHVSPERV 0.002 35 LEEGIGGTI 0.001 32 SQFLEEGIG 0.001 29 WEKSQFLEE 0.000 3 PSIVILGKI 0.000 37 EGIGGTIPH 0.000 28 GWEKSQFLE 0.000 8 LGKIILFLP 0.000 33 QFLEEGIGG 0.000 40 GGTIPHVSP 0.000 39 IGGTIPHVS 0.000 25 IKKGWEKSQ 0.000 30 EKSQFLEEG 0.000 20 RKLKRIKKG 0.000 36 EEGIGGTIP 0.000 22 LKRIKKGWE 0.000 44 PHVSPERVT 0.000

TABLE XII-V7A HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 FLPNGINGI 0.900 4 SLSETFLPN 0.180 1 SPKSLSETF 0.020 6 SETFLPNGI 0.002 3 KSLSETFLP 0.001 7 ETFLPNGIN 0.001 5 LSETFLPNG 0.000 8 TFLPNGING 0.000 2 PKSLSETFL 0.000

TABLE XII-V7B HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 STLGYVALL 0.608 3 NMAYQQSTL 0.600 1 FLNMAYQQS 0.040 7 QQSTLGYVA 0.018 5 AYQQSTLGY 0.008 8 QSTLGYVAL 0.003 6 YQQSTLGYV 0.003 4 MAYQQSTLG 0.001 2 LNMAYQQST 0.001

TABLE XII-V7C HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 167 KLETIILSK 270.000 159 FLGSGTWMK 60.000 175 KLTQEQKSK 30.000 31 GLSEIVLPI 24.300 129 PLWEFLLRL 4.050 109 ALKAANSWR 4.000 148 SLAFTSWSL 1.800 5 ILDLSVEVL 1.800 27 ILRGGLSEI 1.350 165 WMKLETIIL 1.200 128 GPLWEFLLR 1.080 57 AMWTEEAGA 1.000 163 GTWMKLETI 0.675 146 TLSLAFTSW 0.600 131 WEFLLRLLK 0.600 21 KCLGANILR 0.540 12 VLASPAAAW 0.300 185 CMFSLISGS 0.300 13 LASPAAAWK 0.300 37 LPIEWQQDR 0.270 126 GVGPLWEFL 0.270 38 PIEWQQDRK 0.200 134 LLRLLKSQA 0.200 173 LSKLTQEQK 0.100 88 GVVTEDDEA 0.090 69 AQESGIRNK 0.090 7 DLSVEVLAS 0.072 2 SIVILDLSV 0.060 136 RLLKSQAAS 0.060 22 CLGANILRG 0.060 151 FTSWSLGEF 0.045 155 SLGEFLGSG 0.041 181 KSKHCMFSL 0.041 125 NGVGPLWEF 0.030 49 PLSTPPPPA 0.030 4 VILDLSVEV 0.030 145 GTLSLAFTS 0.027 42 QQDRKIPPL 0.027 123 HTNGVGPLW 0.022 51 STPPPPAMW 0.022 133 FLLRLLKSQ 0.022 35 IVLPIEWQQ 0.020 36 VLPIEWQQD 0.020 172 ILSKLTQEQ 0.020 143 ASGTLSLAF 0.020 9 SVEVLASPA 0.020 137 LLKSQAASG 0.020 82 SQIPVVGVV 0.018 179 EQKSKHCMF 0.018 59 WTEEAGATA 0.015 83 QIPVVGVVT 0.015 152 TSWSLGEFL 0.015 176 LTQEQKSKH 0.015 73 GIRNKSSSS 0.012 141 QAASGTLSL 0.012 46 KIPPLSTPP 0.009 11 EVLASPAAA 0.009 103 PESPDRALK 0.009 100 IDPPESPDR 0.006 112 AANSWRNPV 0.006 170 TIILSKLTQ 0.006 120 VLPHTNGVG 0.006 66 TAEAQESGI 0.006 26 NILRGGLSE 0.006 127 VGPLWEFLL 0.005 24 GANILRGGL 0.005 142 AASGTLSLA 0.005 81 SSQIPVVGV 0.005 52 TPPPPAMWT 0.005 3 IVILDLSVE 0.005 171 IILSKLTQE 0.005 119 PVLPHTNGV 0.005 99 SIDPPESPD 0.005 168 LETIILSKL 0.004 17 AAAWKCLGA 0.004 67 AEAQESGIR 0.004 108 RALKAANSW 0.003 15 SPAAAWKCL 0.003 86 VVGVVTEDD 0.003 177 EQEQKSKHC 0.003 14 ASPAAAWKC 0.003 89 VVTEDDEAQ 0.003 154 WSLGEFLGS 0.003 139 KSQAASGTL 0.003 157 GEFLGSGTW 0.003 50 LSTPPPPAM 0.002 34 EIVLPIEWQ 0.002 85 PVVGVVTED 0.002 78 SSSSSQIPV 0.002 182 SKHCMFSLI 0.002 160 LGSGTWMKL 0.002 115 SWRNPVLPH 0.002 33 SEIVLPIEW 0.002 79 SSSSQIPVV 0.002 105 SPDRALKAA 0.002 65 ATAEAQESG 0.002 64 GATAEAQES 0.001 29 RGGLSEIVL 0.001 113 ANSWRNPVL 0.001 140 SQAASGTLS 0.001

TABLE XIII-V1 HLA-A3-10-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 135 SLFPDSLIVK 450.000 281 QLYYGTKYRR 60.000 34 GVIGSGDFAK 40.500 275 LLAAAYQLYY 24.000 294 WLETWLQCRK 20.000 274 GLLAAAYQLY 18.000 17 CLPNGINGIK 9.000 21 GINGIKDARK 9.000 306 GLLSFFFAMV 8.100 271 YLAGLLAAAY 6.000 112 ILIDVSNNMR 6.000 443 ILDLLQLCRY 6.000 227 FLYSFVRDVI 4.500 210 TLWRGPVVVA 4.500 322 CLPMRRSERY 4.000 55 HVVIGSRNPK 3.000 402 ALLISTFHVL 2.700 266 LLSLVYLAGL 2.700 403 LLISTFHVLI 2.700 437 VLPSIVILDL 2.700 404 LISTFHVLIY 2.400 107 LLVGKILIDV 2.025 100 SLWDLRHLLV 2.000 76 VTHHEDALTK 2.000 370 LLSLLAVTSI 1.800 132 YLASLFPDSL 1.800 304 QLGLLSFFFA 1.800 385 ALNWREFSFI 1.800 435 ALVLPSIVIL 1.350 303 KQLGLLSFFF 1.215 307 LLSFFFAMVH 1.200 442 VILDLLQLCR 1.200 298 WLQCRKQLGL 1.200 365 IMSLGLLSLL 0.900 410 VLIYGWKRAF 0.900 140 SLIVKGFNVV 0.900 207 RLFTLWRGPV 0.900 258 TLPIVAITLL 0.900 123 NQYPESNAEY 0.900 278 AAYQLYYGTK 0.900 364 GIMSLGLLSL 0.810 427 YTPPNFVLAL 0.810 220 ISLATEFFLY 0.810 221 SLATFFFLYS 0.720 257 KTLPIVAITL 0.608 333 FLNMAYQQVH 0.600 268 SLVYLAGLLA 0.600 324 PMRRSERYLF 0.600 82 ALTKTNIIFV 0.600 367 SLGLLSLLAV 0.600 203 NLPLRLFTLW 0.600 166 YICSNNIQAR 0.600 219 AISLATFFFL 0.540 147 NVVSAWALQL 0.540 150 SAWALQLGPK 0.450 56 VVIGSRNPKF 0.450 417 RAFEEEYYRF 0.450 45 LTIRLIRCGY 0.450 216 VVVAISLATF 0.450 178 VIELARQLNF 0.400 204 LPLRLFTLWR 0.360 358 EMYISFGIMS 0.360 314 MVHVAYSLCL 0.360 48 RLIRCGYHVV 0.300 317 VAYSLCLPMR 0.300 331 YLFLNMAYQQ 0.300 313 AMVHVAYSLC 0.300 373 LLAVTSIPSV 0.300 269 LVYLAGLLAA 0.300 440 SIVILDLLQL 0.270 222 LATFFFLYSF 0.270 154 LQLGPKDASR 0.270 85 KTNIIFVAIH 0.270 356 RIEMYISFGI 0.270 406 STFHVLIYGW 0.225 396 STLGYVALLI 0.203 432 FVLALVLPSI 0.203 217 VVAISLATFF 0.200 433 VLALVLPSIV 0.200 391 FSFIQSTLGY 0.200 369 GLLSLLAVTS 0.180 224 TFFFLYSFVR 0.180 49 LIRCGYHVVI 0.180 103 DLRHLLVGKI 0.162 111 KILIDVSNNM 0.135 249 KIPIEIVNKT 0.135 264 ITLLSLVYLA 0.135 5 SMMGSPKSLS 0.135 113 LIDVSNNMRI 0.120 262 VAITLLSLVY 0.120 372 SLLAVTSIPS 0.120 397 TLGYVALLIS 0.120 157 GPKDASRQVY 0.120 172 IQARQQVIEL 0.108 243 QQSDFYKIPI 0.108 347 NSWNEEEVWR 0.100 39 GDFAKSLTIR 0.090 218 VAISLATFFF 0.090 384 NALNWREFSF 0.090 285 GTKYRRFPPW 0.090

TABLE XIII-V2 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 22 CLSLPSSWDY 12.000 8 ALSLSLSSGF 2.000 24 SLPSSWDYRC 1.800 5 GLQALSLSLS 0.180 12 SLSSGFTPFS 0.120 10 SLSLSSGFTP 0.060 35 PPCPADFFLY 0.054 11 LSLSSGFTPF 0.045 23 LSLPSSWDYR 0.045 33 CPPPCPADFF 0.045 36 PCPADFFLYF 0.036 32 PCPPPCPADF 0.030 2 GSPGLQALSL 0.027 14 SSGFTPFSCL 0.013 16 GFTPFSCLSL 0.005 13 LSSGFTPFSC 0.005 18 TPFSCLSLPS 0.004 6 LQALSLSLSS 0.002 34 PPPCPADFFL 0.002 17 FTPFSCLSLP 0.002 20 FSCLSLPSSW 0.001 3 SPGLQALSLS 0.001 15 SGFTPFSCLS 0.001 27 SSWDYRCPPP 0.001 21 SCLSLPSSWD 0.000 7 QALSLSLSSG 0.000 9 LSLSLSSGFT 0.000 28 SWDYRCPPPC 0.000 4 PGLQALSLSL 0.000 29 WDYRCPPPCP 0.000 30 DYRCPPPCPA 0.000 1 SGSPGLQALS 0.000 31 YRCPPPCPAD 0.000 26 PSSWDYRCPP 0.000 25 LPSSWDYRCP 0.000 19 PFSCLSLPSS 0.000

TABLE XIII-V5A HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 6 RLFTFWRGPV 0.900 2 NLPLRLFTFW 0.600 3 LPLRLFTFWR 0.540 8 FTFWRGPVVV 0.050 4 PLRLFTFWRG 0.018 1 ENLPLRLFTF 0.012 9 TFWRGPVVVA 0.005 10 FWRGPVVVAI 0.004 7 LFTFWRGPVV 0.000 5 LRLFTFWRGP 0.000

TABLE XIII-V5B HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 20 ELELEFVFLL 4.860 22 ELEFVFLLTL 1.620 18 QTELELEFVF 0.300 5 FSFIQIFCSF 0.225 16 DTQTELELEF 0.060 9 QIFCSFADTQ 0.030 12 CSFADTQTEL 0.015 8 IQIFCSFADT 0.013 23 LEFVFLLTLL 0.013 17 TQTELELEFV 0.013 19 TELELEFVFL 0.012 14 FADTQTELEL 0.012 2 WREFSFIQIF 0.009 3 REFSFIQIFC 0.009 21 LELEFVFLLT 0.006 7 FIQIFCSFAD 0.006 1 NWREFSFIQI 0.005 6 SFIQIFCSFA 0.001 24 EFVFLLTLLL 0.001 11 FCSFADTQTE 0.000 10 IFCSFADTQT 0.000 4 EFSFIQIFCS 0.000 15 ADTQTELELE 0.000 13 SFADTQTELE 0.000

TABLE XIII-V6 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 13 ILFLPCISRK 150.000 2 VLPSIVILGK 90.000 15 FLPCISRKLK 10.000 42 GTIPHVSPER 2.025 12 IILFLPCISR 1.800 6 IVILGKIILF 0.900 7 VILGKIILFL 0.608 35 FLEEGIGGTI 0.405 19 ISRKLKRIKK 0.200 18 CISRKLKRIK 0.200 5 SIVILGKIIL 0.180 8 ILGKIILFLP 0.135 46 HVSPERVTVM 0.090 16 LPCISRKLKR 0.080 23 LKRIKKGWEK 0.060 1 LVLPSIVILG 0.041 39 GIGGTIPHVS 0.027 43 TIPHVSPERV 0.020 22 KLKRIKKGWE 0.018 11 KIILFLPCIS 0.018 10 GKIILFLPCI 0.012 33 SQFLEEGIGG 0.006 3 LPSIVILGKI 0.004 26 IKKGWEKSQF 0.003 25 RIKKGWEKSQ 0.003 27 KKGWEKSQFL 0.002 28 KGWEKSQFLE 0.001 9 LGKIILFLPC 0.001 17 PCISRKLKRI 0.001 29 GWEKSQFLEE 0.000 37 EEGIGGTIPH 0.000 30 WEKSQFLEEG 0.000 21 RKLKRIKKGW 0.000 4 PSIVILGKII 0.000 38 EGIGGTIPHV 0.000 14 LFLPCISRKL 0.000 41 GGTIPHVSPE 0.000 24 KRIKKGWEKS 0.000 31 EKSQFLEEGI 0.000 44 IPHVSPERVT 0.000 34 QFLEEGIGGT 0.000 32 KSQFLEEGIG 0.000 36 LEEGIGGTIP 0.000 45 PHVSPERVTV 0.000 40 IGGTIPHVSP 0.000 20 SRKLKRIKKG 0.000

TABLE XIII-V7A HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 10 FLPNGINGIK 9.000 5 SLSETFLPNG 0.135 1 GSPKSLSETF 0.030 2 SPKSLSETFL 0.006 6 LSETFLPNGI 0.003 8 ETFLPNGING 0.003 4 KSLSETFLPN 0.003 9 TFLPNGINGI 0.002 7 SETFLPNGIN 0.000 3 PKSLSETFLP 0.000

TABLE XIII-V7B HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 MAYQQSTLGY 0.400 2 FLNMAYQQST 0.300 10 STLGYVALLI 0.203 9 QSTLGYVALL 0.027 4 NMAYQQSTLG 0.020 7 YQQSTLGYVA 0.018 8 QQSTLGYVAL 0.018 3 LNMAYQQSTL 0.002 6 AYQQSTLGYV 0.000 1 LFLNMAYQQS 0.000

TABLE XIII-V7C HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 13 VLASPAAAWK 20.000 173 ILSKLTQEQK 20.000 37 VLPIEWQQDR 12.000 168 KLETIILSKL 4.050 127 GVGPLWEFLL 2.430 160 FLGSGTWMKL 1.200 100 SIDPPESPDR 0.600 176 KLTQEQKSKH 0.600 38 LPIEWQQDRK 0.450 164 GTWMKLETII 0.450 134 FLLRLLKSQA 0.300 6 ILDLSVEVLA 0.300 28 ILRGGLSEIV 0.300 5 VILDLSVEVL 0.270 129 GPLWEFLLRL 0.243 167 MKLETIILSK 0.203 32 GLSEIVLPIE 0.203 135 LLRLLKSQAA 0.200 156 SLGEFLGSGT 0.150 58 AMWTEEAGAT 0.150 146 GTLSLAFTSW 0.135 27 NILRGGLSEI 0.135 166 WMKLETIILS 0.120 147 TLSLAFTSWS 0.120 138 LLKSQAASGT 0.100 130 PLWEFLLRLL 0.068 143 AASGTLSLAF 0.060 110 ALKAANSWRN 0.060 109 RALKAANSWR 0.060 66 ATAEAQESGI 0.045 115 NSWRNPVLPH 0.045 159 EFLGSGTWMK 0.041 131 LWEFLLRLLK 0.040 141 SQAASGTLSL 0.036 152 FTSWSLGEFL 0.030 50 PLSTPPPPAM 0.030 137 RLLKSQAASG 0.030 4 IVILDLSVEV 0.030 19 AAWKCLGANI 0.030 125 TNGVGPLWEF 0.027 42 WQQDRKIPPL 0.027 182 KSKHCMFSLI 0.027 31 GGLSEIVLPI 0.024 128 VGPLWEFLLR 0.024 52 STPPPPAMWT 0.022 103 PPESPDRALK 0.020 10 SVEVLASPAA 0.020 149 SLAFTSWSLG 0.020 121 VLPHTNGVGP 0.020 112 KAANSWRNPV 0.018 175 SKLTQEQKSK 0.015 148 LSLAFTSWSL 0.013 12 EVLASPAAAW 0.013 8 DLSVEVLASP 0.013 69 EAQESGIRNK 0.013 74 GIRNKSSSSS 0.012 67 TAEAQESGIR 0.012 83 SQIPVVGVVT 0.010 89 GVVTEDDEAQ 0.009 47 KIPPLSTPPP 0.009 14 LASPAAAWKC 0.009 158 GEFLGSGTWM 0.009 21 WKCLGANILR 0.008 177 LTQEQKSKHC 0.007 179 QEQKSKHCMF 0.006 113 AANSWRNPVL 0.006 84 QIPVVGVVTE 0.006 178 TQEQKSKHCM 0.006 150 LAFTSWSLGE 0.006 78 KSSSSSQIPV 0.006 122 LPHTNGVGPL 0.005 3 SIVILDLSVE 0.005 36 IVLPIEWQQD 0.005 23 CLGANILRGG 0.005 171 TIILSKLTQE 0.005 81 SSSQIPVVGV 0.005 22 KCLGANILRG 0.004 35 EIVLPIEWQQ 0.004 119 NPVLPHTNGV 0.003 162 GSGTWMKLET 0.003 142 QAASGTLSLA 0.003 124 HTNGVGPLWE 0.003 90 VVTEDDEAQD 0.003 172 IILSKLTQEQ 0.003 181 QKSKHCMFSL 0.003 9 LSVEVLASPA 0.002 51 LSTPPPPAMW 0.002 87 VVGVVTEDDE 0.002 33 LSEIVLPIEW 0.002 91 VTEDDEAQDS 0.002 165 TWMKLETIIL 0.002 29 LRGGLSEIVL 0.002 70 AQESGIRNKS 0.002 132 WEFLLRLLKS 0.002 43 QQDRKIPPLS 0.002 92 TEDDEAQDSI 0.002 79 SSSSSQIPVV 0.002 60 WTEEAGATAE 0.002 153 TSWSLGEFLG 0.002 15 ASPAAAWKCL 0.002

TABLE XIV-V1 HLA-A1101 -9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 56 VVIGSRNPK 3.000 409 HVLIYGWKR 1.200 249 KIPIEIVNK 1.200 35 VIGSGDFAK 1.200 175 ROOVIELAR 0.720 417 RAFEEEYYR 0.480 3 SISMMGSPK 0.400 279 AYQLYYGTK 0.400 136 LFPDSLIVK 0.400 381 SVSNALNWR 0.400 241 RNQQSDFYK 0.360 282 LYYGTKYRR 0.320 225 FFFLYSFVR 0.240 21 GINGIKDAR 0.240 53 GYHVVIGSR 0.240 87 NIIFVAIHR 0.240 18 LPNGINGIK 0.200 443 ILDLLQLCR 0.160 103 DLRHLLVGK 0.120 34 GVIGSGDFA 0.090 322 CLPMRRSER 0.080 113 LIDVSNNMR 0.080 155 QLGPKDASR 0.080 318 AYSLCLPMR 0.080 269 LVYLAGLLA 0.080 281 QLYYGTKYR 0.080 294 WLETWLQCR 0.080 97 HYTSLWDLR 0.080 295 LETWLQCRK 0.060 441 IVILDLLQL 0.060 306 GLLSFFFAM 0.054 199 REIENLPLR 0.054 22 INGIKDARK 0.040 148 VVSAWALQL 0.040 77 THHEDALTK 0.040 108 LVGKILIDV 0.040 223 ATFFFLYSF 0.040 261 IVAITLLSL 0.040 167 ICSNNIQAR 0.040 164 QVYICSNNI 0.040 43 KSLTIRLIR 0.036 233 RDVIHPYAR 0.036 48 RLIRCGYHV 0.036 274 GLLAAAYQL 0.036 330 RYLFLNMAY 0.036 408 FHVLIYGWK 0.030 85 KTNIIFVAI 0.030 436 LVLPSIVIL 0.030 303 KQLGLLSFF 0.027 353 EVWRIEMYI 0.024 191 DLGSLSSAR 0.024 254 IVNKTLPIV 0.020 90 FVAIHREHY 0.020 151 AWALQLGPK 0.020 83 LTKTNIIFV 0.020 98 YTSLWDLRH 0.020 231 FVRDVIHPY 0.020 400 YVALLISTF 0.020 217 VVAISLAFT 0.020 402 ALLISTFHV 0.018 64 KFASEFFPH 0.018 140 SLIVKGFNV 0.018 214 GPVVVAISL 0.018 135 SLFPDSLIV 0.016 205 PLRLFTLWR 0.016 209 FLTWRGPVV 0.015 264 ITLLSLVYL 0.015 396 STLGYVALL 0.015 319 YSLCLPMRR 0.012 394 IQSTLGYVA 0.012 30 KVTVGVIGS 0.012 270 VYLAGLLAA 0.012 203 NLPLRLFTL 0.012 425 RFYTPPNFV 0.012 242 NQQSDFYKI 0.012 287 KYRRFPPWL 0.012 435 ALVLPSIVI 0.012 265 TLLSLVYLA 0.012 299 LQCRKQLGL 0.012 313 AMVHVAYSL 0.012 40 DFAKSLTIR 0.012 106 HLLVGKILI 0.012 426 FYTPPNFVL 0.012 385 ALNWREFSF 0.012 219 AISLATFFF 0.012 304 QLGLLSFFF 0.012 221 SLATFFFLY 0.012 427 YTPPNFVLA 0.010 285 GTKYRRFPP 0.009 280 YQLYYGTKY 0.009 397 TLGYVALLI 0.008 367 SLGLLSLLA 0.008 166 YICSNNIQA 0.008 258 TLPIVAITL 0.008 317 VAYSLCLPM 0.008 100 SLWDLRHLL 0.008 210 TLWRGPVVV 0.008 365 IMSLGLLSL 0.008 263 AITLLSLVY 0.008 360 YISFGIMSL 0.008

TABLE XIV-V2 HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 24 SLPSSWDYR 0.080 5 GLQALSLSL 0.024 17 FTPFSCLSL 0.020 3 SPGLQALSL 0.004 12 SLSSGFTPF 0.004 37 CPADFFLYF 0.004 21 SCLSLPSSW 0.003 33 CPPPCPADF 0.002 23 LSLPSSWDY 0.001 6 LQALSLSLS 0.001 16 GFTPFSCLS 0.001 32 RCPPPCPAD 0.001 36 PCPADFFLY 0.001 35 PPCPADFFL 0.001 7 QALSLSLSS 0.001 10 SLSLSSGFT 0.000 15 SGFTPFSCL 0.000 22 CLSLPSSWD 0.000 8 ALSLSLSSG 0.000 18 TPFSCLSLP 0.000 25 LPSSWDYRC 0.000 9 LSLSLSSGF 0.000 1 SGSPGLQAL 0.000 31 YRCPPPCPA 0.000 34 PPPCPADFF 0.000 30 DYRCPPPCP 0.000 11 LSLSSGFTP 0.000 14 SSGFTPFSC 0.000 2 GSPGLQALS 0.000 19 PFSCLSLPS 0.000 29 WDYRCPPPC 0.000 27 SSWDYRCPP 0.000 13 LSSGFTPFS 0.000 20 FSCLSLPSS 0.000 28 SWDYRCPPP 0.000 4 PGLQALSLS 0.000 26 PSSWDYRCP 0.000

TABLE XIV-V5 HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 3 PLRLFTFWR 0.024 7 FTFWRGPVV 0.020 1 NLPLRLFTF 0.012 8 TFWRGPVVV 0.004 2 LPLRLFTFW 0.003 6 LFTFWRGPV 0.002 5 RLFTFWRGP 0.000 9 FWRGPVVVA 0.000 4 LRLFTFWRG 0.000

TABLE XIV-V5B HLA-A11-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 24 FVFLLTLLL 0.080 16 TQTELELEF 0.012 17 QTELELEFV 0.010 6 FIQIFCSFA 0.004 2 REFSFIQIF 0.004 5 SFIQIFCSF 0.003 7 IQIFCSFAD 0.003 18 TELELEFVF 0.003 20 LELEFVFLL 0.003 22 LEFVFLLTL 0.002 12 SFADTQTEL 0.002 19 ELELEFVFL 0.001 23 EFVFLLTLL 0.001 8 QIFCSFADT 0.001 14 ADTQTELEL 0.000 1 WREFSFIQI 0.000 15 DTQTELELE 0.000 21 ELEFVFLLT 0.000 9 IFCSFADTQ 0.000 13 FADTQTELE 0.000 10 FCSFADTQT 0.000 4 FSFIQIFCS 0.000 3 EFSFIQIFC 0.000 11 CSFADTQTE 0.000

TABLE XIV-V6 HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 2 LPSIVILGK 0.400 12 ILFLPCISR 0.320 13 LFLPCISRK 0.300 23 KRIKKGWEK 0.180 15 LPCISRKLK 0.100 42 TIPHVSPER 0.080 5 IVILGKIIL 0.060 19 SRKLKRIKK 0.040 45 HVSPERVTV 0.020 10 KIILFLPCI 0.018 16 PCISRKLKR 0.012 6 VILGKIILF 0.012 38 GIGGTIPHV 0.012 7 ILGKIILFL 0.008 21 KLKRIKKGW 0.006 41 GTIPHVSPE 0.005 4 SIVILGKII 0.003 18 ISRKLKRIK 0.002 17 CISRKLKRI 0.002 43 IPHVSPERV 0.002 32 SQFLEEGIG 0.001 24 RIKKGWEKS 0.001 27 KGWEKSQFL 0.001 1 VLPSIVILG 0.001 26 KKGWEKSQF 0.001 11 IILFLPCIS 0.001 33 QFLEEGIGG 0.001 31 KSQFLEEGI 0.001 35 LEEGIGGTI 0.001 14 FLPCISRKL 0.000 34 FLEEGIGGT 0.000 46 VSPERVTVM 0.000 9 GKIILFLPC 0.000 28 GWEKSQFLE 0.000 37 EGIGGTIPH 0.000 29 WEKSQFLEE 0.000 8 LGKIILFLP 0.000 40 GGTIPHVSP 0.000 20 RKLKRIKKG 0.000 3 PSIVILGKI 0.000 22 LRKIKKGWE 0.000 39 IGGTIPHVS 0.000 36 EEGIGGTIP 0.000 25 IKKGWEKSQ 0.000 30 EKSQFLEEG 0.000 44 PHVSPERVT 0.000

TABLE XIV-V7A HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 FLPNGINGI 0.004 1 SPKSLSETF 0.002 4 SLSETFLPN 0.001 7 ETFLPNGIN 0.001 8 TFLPNGING 0.001 6 SETFLPNGI 0.001 3 KSLSETFLP 0.000 2 PKSLSETFL 0.000 5 LSETFLPNG 0.000

TABLE XIV-V7B HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 STLGYVALL 0.015 7 QQSTLGYVA 0.012 5 AYQQSTLGY 0.008 6 YQQSTLGYV 0.006 3 NMAYQQSTL 0.004 4 MAYQQSTLG 0.000 1 FLNMAYQQS 0.000 8 QSTLGYVAL 0.000 2 LNMAYQQST 0.000

TABLE XIV-V7C HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 167 KLETIILSK 2.400 159 FLGSGPNMK 0.800 175 KLTQEQKSK 0.600 21 KCLGANILR 0.360 128 GPLWEFLLR 0.360 131 WEFLLRLLK 0.240 13 LASPAAAWK 0.200 88 GVVTEDDEA 0.090 109 ALKAANSWR 0.080 69 AQESGIRNK 0.060 163 GTWMKLETI 0.060 37 LPIEWQQDR 0.060 126 GVGPLWEFL 0.060 38 PIEWQQDRK 0.040 31 GLSEIVLPI 0.024 173 LSKLTQEQK 0.020 9 SVEVLASPA 0.020 145 GTLSLAFTS 0.013 2 SIVILDLSV 0.012 67 AEAQESGIR 0.012 151 FTSWSLGEF 0.010 176 LTQEQKSKH 0.010 51 STPPPPAMW 0.010 59 WTEEAGATA 0.010 123 HTNGVGPLW 0.010 11 EVLASPAAA 0.009 82 SQIPVVGVV 0.009 108 RALKAANSW 0.009 57 AMWTEEAGA 0.008 165 WMKLETIIL 0.008 148 SLAFTSWSL 0.008 4 VILDLSVEV 0.006 103 PESPDRALK 0.006 42 QQDRKIPPL 0.006 24 GANILRGGL 0.006 35 IVLPIEWQQ 0.006 5 ILDLSVEVL 0.004 134 LLRLLKSQA 0.004 100 IDPPESPDR 0.004 141 QAASGTLSL 0.004 27 ILRGGLSEI 0.004 146 TLSLAFTSW 0.004 12 VLASPAAAW 0.004 17 AAAWKCLGA 0.004 157 GEFLGSGTW 0.004 3 IVILDLSVE 0.003 119 PVLPHTNGV 0.003 89 VVTEDDEAQ 0.002 66 TAEAQESGI 0.002 86 VVGVVTEDD 0.002 142 AASGTLSLA 0.002 112 AANSWRNPV 0.002 181 KSKHCMFSL 0.002 136 RLLKSQAAS 0.002 179 EQKSKHCMF 0.002 33 SEIVLPIEW 0.002 129 PLWEFLLRL 0.002 170 TIILSKLTQ 0.001 73 GIRNKSSSS 0.001 29 RGGLSEIVL 0.001 46 KIPPLSTPP 0.001 41 WQQDRKIPP 0.001 26 NILRGGLSE 0.001 105 SPDRALKAA 0.001 65 ATAEAQESG 0.001 15 SPAAAWKCL 0.001 90 VTEDDEAQD 0.001 158 EFLGSGTWM 0.001 10 VEVLASPAA 0.001 22 CLGANILRG 0.001 185 CMFSLISGS 0.001 184 HCMFSLISG 0.001 127 VGPLWEFLL 0.001 139 KSQAASGTL 0.001 125 NGVGPLWEF 0.001 64 GATAEAGES 0.001 140 SQAASGTLS 0.001 171 IILSKLTQE 0.001 96 AQDSIDPPE 0.001 168 LETIILSKL 0.001 178 QEQKSKHCM 0.001 101 DPPESPDRA 0.001 164 TWMKLETII 0.000 49 PLSTPPPPA 0.000 18 AAWKCLGAN 0.000 143 ASGTLSLAF 0.000 150 AFTSWSLGE 0.000 36 VLPIEWQQD 0.000 83 QIPVVGVVT 0.000 160 LGSGTWMKL 0.000 52 TPPPPAMWT 0.000 172 ILSKLTQEQ 0.000 115 SWRNPVLPH 0.000 99 SIDPPESPD 0.000 149 LAFTSWSLG 0.000 113 ANSWRNPVL 0.000 155 SLGEFLGSG 0.000 137 LLKSQAASG 0.000 120 VLPHTNGVG 0.000 78 SSSSSQIPV 0.000

TABLE XV-V1 HLA-A1101-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 34 GVIGSGDFAK 27.000 55 HVVIGSRNPK 3.000 76 VTHHEDALTK 2.000 135 SLFPDSLIVK 1.600 21 GINGIKDARK 1.200 294 WLETWLQCRK 0.400 278 AAYQLYYGTK 0.400 150 SAWALQLGPK 0.400 17 CLPNGINGIK 0.400 281 QLYYGTKYRR 0.320 442 VILDLLQLCR 0.240 24 TFFFLYSFVR 0.240 407 TFHVLIYGWK 0.200 154 LQLGPKDASR 0.180 318 AYSLCLPMRR 0.160 204 LPLRLFTLWR 0.120 112 ILIDVSNNMR 0.120 280 YQLYYGTKYR 0.090 257 KTLPIVAITL 0.090 303 KQLGLLSFFF 0.081 166 YICSNNIQAR 0.080 269 LVYLAGLLAA 0.080 317 VAYSLCLPMR 0.080 240 ARNQQSDFYK 0.060 321 LCLPMRRSER 0.060 147 NVVSAWALQL 0.060 183 RQLNFIPIDL 0.054 364 GIMSLGLLSL 0.048 406 STFHVLIYGW 0.040 254 IVNKTLPIVA 0.040 314 MVHVAYSLCL 0.040 316 HVAYSLCLPM 0.040 356 PIEMYISFGI 0.036 425 RFYTPPNFVL 0.036 102 WDLRHLLVGK 0.030 248 YKIPIEIVNK 0.030 56 VVIGSRNPKF 0.030 285 GTKYRRFPPW 0.030 216 VVVAISLATF 0.030 83 LTKTNIIFVA 0.030 85 KTNIIFVAIH 0.030 396 STLGYVALLI 0.030 432 FVLALVLPSI 0.030 264 ITLLSLVYLA 0.030 163 RQVYICSNNI 0.027 416 KRAFEEEYYR 0.024 86 TNIIFVAIHR 0.024 39 GDFAKSLTIR 0.024 417 RAFEEEYYRF 0.024 207 RLFTLWRGPV 0.024 217 VVAISLATFF 0.020 223 ATFFFLYSFV 0.020 400 YVALLISTFH 0.020 261 IVAITLLSLV 0.020 32 TVGVIGSGDF 0.020 142 IVKGFNVVSA 0.020 231 FVRDVIHPYA 0.020 73 VVDVTHHEDA 0.020 340 QVHANIENSW 0.020 427 YTPPNFVLAL 0.020 399 GYVALLISTF 0.018 111 KILIDVSNNM 0.018 274 GLLAAAYQLY 0.018 48 RLIRCGYHVV 0.018 306 GLLSFFFAMV 0.018 100 SLWDLRHLLV 0.016 45 LTIRLIRCGY 0.015 209 FTLWRGPVVV 0.015 409 HVLIYGWKRA 0.015 408 FHVLIYGWKR 0.012 243 QQSDFYKIPI 0.012 440 SIVILDLLQL 0.012 24 GIDKARKVTV 0.012 304 QLGLLSFFFA 0.012 145 GFNVVSAWAL 0.012 359 MYISFGIMSL 0.012 172 IQARQQVIEL 0.012 121 RINQYPESNA 0.012 123 NQYPESNAEY 0.012 165 VYICSNNIQA 0.012 107 LLVGKILIDV 0.012 219 AISLATFFFL 0.012 268 SLVYLAGLLA 0.012 376 VTSIPSVSNA 0.010 2 ESISMMGSPK 0.009 401 VALLISTFHV 0.009 214 GPVVVAISLA 0.009 218 VAISLATFFF 0.009 384 NALNWREFSF 0.009 367 SLGLLSLLAV 0.008 307 LLSFFFAMVH 0.008 437 VLPSIVILDL 0.008 227 FLYSFVRDVI 0.008 42 AKSLTIRLIR 0.008 113 LIDVSNNMRI 0.008 210 TLWRGPVVVA 0.008 178 VIELARQLNF 0.008 298 WLQCRKQLGL 0.008 404 LISTFHVLIY 0.008 82 ALTKTNIIFV 0.008

TABLE XV-2V HLA-A1101-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 16 GFTPFSCLSL 0.012 22 CLSLPSSWDY 0.008 23 LSLPSSWDYR 0.006 32 RCPPPCPADF 0.006 8 ALSLSLSSGF 0.004 33 CPPPCPADFF 0.002 6 LQALSLSLSS 0.001 2 GSPGLQALSL 0.001 5 GLQALSLSLS 0.001 10 SLSLSSGFTP 0.001 30 DYRCPPPCPA 0.001 17 FTPFSCLSLP 0.001 18 TPFSCLSLPS 0.001 24 SLPSSWDYRC 0.001 35 PPCPADFFLY 0.001 34 PPPCPADFFL 0.001 36 PCPADFFLYF 0.000 12 SLSSGFTPFS 0.000 11 LSLSSGFTPF 0.000 21 SCLSLPSSWD 0.000 7 QALSLSLSSG 0.000 3 SPGLQALSLS 0.000 20 FSCLSLPSSW 0.000 14 SSGFTPFSCL 0.000 4 PGLQALSLSL 0.000 13 LSSGFTPFSC 0.000 29 WDYRCPPPCP 0.000 27 SSWDYRCPPP 0.000 15 SGFTPFSCLS 0.000 9 LSLSLSSGFT 0.000 31 YRCPPPCPAD 0.000 19 PFSCLSLPSS 0.000 25 LPSSWDYRCP 0.000 28 SWDYRCPPPC 0.000 1 SGSPGLQALS 0.000 26 PSSWDYRCPP 0.000

TABLE XV-V5A HLA-A1101-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 3 LPLRLFTFWR 0.180 6 RLFTFWRGPV 0.024 8 FTFWRGPVVV 0.020 9 TFWRGPVVVA 0.004 2 NLPLRLFTFW 0.004 7 LTFTWRGPVV 0.002 1 ENLPLRLFTF 0.001 10 FWRGPVVVAI 0.000 4 PLRLFTFWRG 0.000 5 LRLFTFWRGP 0.000

TABLE XV-V5B HLA-A1101-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 18 QTELELEFVF 0.030 16 DTQTELELEF 0.006 17 TQTELELEFV 0.006 14 FADTQTELEL 0.004 20 ELELEEVELL 0.004 6 SFIQIFCSFA 0.003 22 ELEFVFLLTL 0.002 24 EFVFLLTLLL 0.002 7 FIQIFCSFAD 0.001 23 LEFVFLLTLL 0.001 8 IQIFCSFADT 0.001 19 TELELEFVFL 0.001 9 QIFCSFADTQ 0.001 3 REFSFIQIFC 0.001 1 NWREFSFIQI 0.000 12 CSFADTQTEL 0.000 5 FSFIQIFCSF 0.000 13 SFADTQTELE 0.000 2 WREFSFIQIF 0.000 11 FCSFADTQTE 0.000 10 IFCSFADTQT 0.000 4 EFSFIQIFCS 0.000 21 LELEFVFLLT 0.000 15 ADTQTELELE 0.000

TABLE XV-V6 HLA-A1101-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 42 GTIPHVSPER 0.900 2 VLPSIVILGK 0.800 13 ILFLPCISRK 0.800 12 IILFLPCISR 0.240 15 FLPCISRKLK 0.200 16 LPCISRKLKR 0.080 6 IVILGKIILF 0.060 19 ISRKLKRIKK 0.040 18 CISRKLKRIK 0.040 23 LKRIKKGWEK 0.040 46 HVSPERVTVM 0.020 5 SIVILGKIIL 0.012 7 VILGKIILFL 0.012 1 LVLPSIVILG 0.006 35 FLEEGIGGTI 0.004 43 TIPHVSPERV 0.004 33 SQFLEEGIGG 0.002 3 LPSIVILGKI 0.002 11 KIILFLPCIS 0.002 39 GIGGTIPHVS 0.001 8 ILGKIILFLP 0.001 22 KLKRIKKGWE 0.001 10 GKIILFLPCI 0.001 25 RIKKGWEKSQ 0.001 27 KKGWEKSQFL 0.001 21 RKLKRIKKGW 0.000 28 KGWEKSQFLE 0.000 37 EEGIGGTIPH 0.000 14 LFLPCISRKL 0.000 34 QFLEEGIGGT 0.000 26 IKKGWEKSQF 0.000 17 PCISRKLKRI 0.000 29 GWEKSQFLEE 0.000 24 KRIKKGWEKS 0.000 38 EGIGGTIPHV 0.000 41 GGTIPHVSPE 0.000 32 KSQFLEEGIG 0.000 36 LEEGIGGTIP 0.000 31 EKSQFLEEGI 0.000 30 WEKSQFLEEG 0.000 9 LGKIILFLPC 0.000 45 PHVSPERVTV 0.000 44 IPHVSPERVT 0.000 40 IGGTIPHVSP 0.000 4 PSIVILGLII 0.000 20 SRKLKRIKKG 0.000

TABLE XV-V7A HLA-A1101-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 10 FLPNGINGIK 0.400 9 TFLPNGINGI 0.003 2 SPKSLSETFL 0.002 8 ETFLPNGING 0.001 1 GSPKSLSETF 0.001 5 SLSETFLPNG 0.000 6 LSETFLPNGI 0.000 4 KSLSETFLPN 0.000 7 SETFLPNGIN 0.000 3 PKSLSETFLP 0.000

TABLE XV-V7B A1101-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 10 STLGYVALLI 0.030 7 YQQSTLGYVA 0.012 5 MAYQQSTLGY 0.008 8 QQSTLGYVAL 0.006 6 AYQQSTLGYV 0.004 3 LNMAYQQSTL 0.001 2 FLNMAYQQST 0.000 4 NMAYQQSTLG 0.000 1 LFLNMAYQQS 0.000 9 QSTLGYVALL 0.000

TABLE XV-V7C A1101-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 173 ILSKLTQEQK 0.400 13 VLASPAAAWK 0.400 38 LPIEWQQDRK 0.300 109 RALKAANSWR 0.180 127 GVGPLWEFLL 0.180 159 EFLGSGTWMK 0.180 100 SIDPPESPDR 0.080 37 VLPIEWQQDR 0.080 167 MKLETIILSK 0.060 164 GTWMKLETII 0.060 146 GTLSLAFTSW 0.045 131 LWEFLLRLLK 0.040 67 TAEAQESGIR 0.040 4 IVILDLSVEV 0.030 103 PPESPDRALK 0.020 10 SVEVLASPAA 0.020 129 GPLWEFLLRL 0.018 175 SKLTQEQKSK 0.015 176 KLTQEQKSKH 0.012 141 SQAASGTLSL 0.012 168 KLETIILSKL 0.012 66 ATAEAQESGI 0.010 152 FTSWSLGEFL 0.010 12 EVLASPAAAW 0.009 89 GVVTEDDEAQ 0.009 160 FLGSGTWMKL 0.008 21 WKCLGANILR 0.008 128 VGPLWEFLLR 0.008 5 VILDLSVEVL 0.006 112 KAANSWRNPV 0.006 134 FLLRLLKSQA 0.006 69 EAQESGIRNK 0.006 27 NILRGGLSEI 0.006 178 TQEQKSKHCM 0.006 42 WQQDRKIPPL 0.006 6 ILDLSVEVLA 0.004 135 LLRLLKSQAA 0.004 143 AASGTLSLAF 0.004 19 AAWKCLGANI 0.004 28 ILRGGLSEIV 0.004 158 GEFLGGSTWM 0.004 36 IVLPIEWQQD 0.003 119 NPVLPHTNGV 0.003 124 HTNGVGPLWE 0.002 113 AANSWRNPVL 0.002 122 LPHTNGVGPL 0.002 52 STPPPPAMWT 0.002 90 VVTEDDEAQD 0.002 142 QAASGTLSLA 0.002 87 VVGVVTEDDE 0.002 151 AFTSWSLGEF 0.002 31 GGLSEIVLPI 0.002 137 RLLKSQAASG 0.002 22 KCLGANILRG 0.002 74 GIRNKSSSSS 0.001 47 KIPPLSTPPP 0.001 32 GLSEIVLPIE 0.001 76 RNKSSSSSQI 0.001 78 KSSSSSQIPV 0.001 60 WTEEAGATAE 0.001 91 VTEDDEAQDS 0.001 83 SQIPVVGVVT 0.001 170 ETIILSKLTQ 0.001 11 VEVLASPAAA 0.001 165 TWMKLETIIL 0.001 125 TNGVGPLWEF 0.001 110 ALKAANSWRN 0.001 166 WMKLETIILS 0.001 115 NSWRNPVLPH 0.001 150 LAFTSWSLGE 0.001 58 AMWTEEAGAT 0.001 3 SIVILDLSVE 0.001 182 KSKHCMFSLI 0.001 171 TIILSKLTQE 0.001 70 AQESGIRNKS 0.001 181 QKSKHCMFSL 0.001 172 IILSKLTQEQ 0.001 148 LSLAFTSWSL 0.001 65 GATAEAQESG 0.001 25 GANILRGGLS 0.001 97 AQDSIDPPES 0.001 43 QQDRKIPPLS 0.001 92 TEDDEAQDSI 0.001 61 TEEAGATAEA 0.001 179 QEQKSKHCMF 0.001 177 LTQEQKSKHC 0.001 147 TLSLAFTSWS 0.000 121 VLPHTNGVGP 0.000 17 PAAAMKCLGA 0.000 138 LLKSQAASGT 0.000 29 LRGGLSEIVL 0.000 53 TPPPPAMWTE 0.000 14 LASPMAAAKC 0.000 84 QIPVVGVVTE 0.000 50 PLSTPPPPAM 0.000 185 HCMFSLISGS 0.000 57 PAMWTEEAGA 0.000 149 SLAFTSWSLG 0.000 33 LSEIVLPIEW 0.000 156 SLGEFLGSGT 0.000

TABLE XVI-V1 HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 287 KYRRFPPWL 400.000 426 FYTPPNFVL 240.000 337 AYQQVHANI 105.000 283 YYGTKYRRF 100.000 228 LYSFVRDVI 70.000 390 EFSFIQSTL 28.000 362 SFGIMSLGL 20.000 418 AFEEEYYRF 18.000 330 RYLFLNMAY 18.000 378 SIPSVSNAL 10.080 124 QYPESNAEY 9.900 399 GYVALLIST 9.000 177 QVIELARQL 8.640 184 QLNFIPIDL 8.400 258 TLPIVAITL 8.400 313 AMVHVAYSL 8.400 214 GPVVVAISL 8.400 246 DFYKIPIEI 7.700 270 VYLAGLLAA 7.500 359 MYISFGIMS 7.500 268 SLVYLAGLL 7.200 291 FPPWLETWL 7.200 366 MSLGLLSLL 7.200 220 ISLATFFFL 7.200 403 LLISTFHVL 7.200 303 KQLGLLSFF 7.200 436 LVLPSIVIL 7.200 200 EIENLPLRL 7.200 61 RNPKFASEF 6.600 428 TPPNFVLAL 6.000 274 GLLAAAYQL 6.000 125 YPESNAEYL 6.000 363 FGIMSLGLL 6.000 264 ITLLSLVYL 6.000 396 STLGYVALL 6.000 297 TWLQCRKQL 6.000 259 LPIVAITLL 6.000 5 SMMGSPKSL 6.000 203 NLPLRLFTL 6.000 441 IVILDLLQL 6.000 187 FIPIDLGSL 6.000 146 FNVVSAWAL 6.000 267 LSLYVLAGL 6.000 99 TSLWDLRHL 6.000 100 SLWDLRHLL 5.760 438 LPSIVILDL 5.600 85 KTNIIFVAI 5.040 247 FYKIPIEIV 5.000 423 YYRFYTPPN 5.000 128 SNAEYLASL 4.800 41 FAKSLTIRL 4.800 37 GSGDFAKSL 4.800 173 QARQQVIEL 4.400 300 QCRKQLGLL 4.000 75 DVTHHEDAL 4.000 395 QSTLGYVAL 4.000 299 LQCRKQLGL 4.000 133 LASLFPDSL 4.000 365 IMSLGLLSL 4.000 148 VVSAWALQL 4.000 360 YISFGIMSL 4.000 261 IVAITLLSL 4.000 196 SSAREIENL 4.000 129 NAEYLASLF 3.600 218 VAISLATFF 3.600 385 ALNWREFSF 3.000 33 VGVIGSGDF 3.000 400 YVALLISTF 2.400 304 QLGLLSFFF 2.400 383 SNALNWREF 2.200 57 VIGSRNPKF 2.200 223 ATFFFLYSF 2.000 411 LIYGWKRAF 2.000 219 AISLATFFF 2.000 62 NPKFASEFF 2.000 82 ALTKTNIIF 2.000 239 YARNQQSDF 2.000 217 VVAISLATF 2.000 242 NQQSDFYKI 1.980 81 DALTKTNII 1.800 17 CLPNGINGI 1.800 349 WNEEEVWRI 1.800 171 NIQARQQVI 1.800 290 RFPPWLETW 1.800 105 RHLLVGKIL 1.680 193 GSLSSAREI 1.650 112 ILIDVSNNM 1.512 435 ALVLPSIVI 1.500 106 HLLVGKILI 1.500 134 ASLFPDSLI 1.500 253 EIVNKTLPI 1.500 371 LSLLAVTSI 1.500 353 EVWRIEMYI 1.400 397 TLGYVALLI 1.400 433 VLALVLPSI 1.400 186 NFIPIDLGS 1.260 164 QVYICSNNI 1.200 180 ELARQLNFI 1.200 425 RFYTPPNFV 1.200 386 LNWREFSFI 1.200

TABLE XVI-V2 HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 5 GLQALSLSL 7.200 17 FTPFSCLSL 6.000 1 SGSPGLQAL 5.760 15 SGFTPFSCL 4.800 3 SPGLQALSL 4.000 33 CPPPCPADF 3.600 9 LSLSLSSGF 3.600 37 CPADFFLYF 2.880 12 SLSSGFTPF 2.400 16 GFTPFSCLS 0.600 30 DYRCPPPCP 0.500 35 PPCPADFFL 0.480 34 PPPCPADFF 0.300 23 LSLPSSWDY 0.180 2 GSPGLQALS 0.180 21 SCLSLPSSW 0.180 7 QALSLSLSS 0.180 14 SSGFTPFSC 0.100 10 SLSLSSGFT 0.100 6 LQALSLSLS 0.100 25 LPSSWDYRC 0.100 13 LSSGFTPFS 0.100 20 FSCLSLPSS 0.100 19 PFSCLSLPS 0.060 32 RCPPPCPAD 0.036 36 PCPADFFLY 0.018 24 SLPSSWDYR 0.015 4 PGLQALSLS 0.015 11 LSLSSGFTP 0.015 27 SSWDYRCPP 0.012 31 YRCPPPCPA 0.012 18 TPFSCLSLP 0.010 29 WDYRCPPPC 0.010 8 ALSLSLSSG 0.010 28 SWDYRCPPP 0.010 22 CLSLPSSWD 0.010 26 PSSWDYRCP 0.001

TABLE XVI-V5A HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 NLPLRLFTF 3.000 8 TFWRGPVVV 0.500 6 LFTFWRGPV 0.500 2 LPLRLFTFW 0.216 7 FTFWRGPVV 0.100 9 FWRGPVVVA 0.100 5 RLFTFWRGP 0.020 4 LRLFTFWRG 0.002 3 PLRLFTFWR 0.001

TABLE XVI-V5B HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 23 EFVFLLTLL 36.000 12 SFADTQTEL 26.400 5 SFIQIFCSF 25.200 19 ELELEFVFL 7.200 24 FVFLLTLLL 4.800 16 TQTELELEF 3.168 20 LELEFVFLL 0.720 3 EFSFIQIFC 0.700 2 REFSFIQIF 0.480 14 ADTQTELEL 0.440 18 TELELEFVF 0.432 22 LEFVFLLTL 0.400 21 ELEFVFLLT 0.252 1 WREFSFIQI 0.180 6 FIQIFCSFA 0.150 17 QTELELEFV 0.150 8 QIFCSFADT 0.120 10 FCSFADTQT 0.100 4 FSFIQIFCS 0.100 9 IFCSFADTQ 0.050 7 IQIFCSFAD 0.015 15 DTQTELELE 0.015 11 CSFADTQTE 0.012 13 FADTQTELE 0.010

TABLE XVI-V6 HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 27 KGWEKSQFL 11.520 14 FLPCISRKL 9.240 5 IVILGKIIL 6.000 7 ILGKIILFL 5.600 31 KSQFLEEGI 3.600 10 KIILFLPCI 3.000 6 VILGKIILF 3.000 4 SIVILGKII 1.800 17 CISRKLKRI 1.000 46 VSPERVTVM 0.900 26 KKGWEKSQF 0.400 21 KLKRIKKGW 0.280 3 PSIVILGKI 0.231 24 RIKKGWEKS 0.220 35 LEEGIGGTI 0.210 34 FLEEGIGGT 0.180 11 IILFLPCIS 0.180 39 IGGTIPHVS 0.140 45 HVSPERVTV 0.120 38 GIGGTIPHV 0.100 43 IPHVSPERV 0.100 33 QFLEEGIGG 0.090 13 LFLPCISRK 0.090 42 TIPHVSPER 0.023 9 GKIILFLPC 0.022 1 VLPSIVILG 0.021 41 GTIPHVSPE 0.018 28 GWEKSQFLE 0.015 37 EGIGGTIPH 0.015 2 LPSIVILGK 0.014 8 LGKIILFLP 0.014 18 ISRKLKRIK 0.012 32 SQFLEEGIG 0.010 40 GGTIPHVSP 0.010 15 LPCISRKLK 0.010 12 ILFLPCISR 0.010 23 KRIKKGWEK 0.003 20 RKLKRIKKG 0.003 16 PCISRKLKR 0.002 44 PHVSPERVT 0.002 29 WEKSQFLEE 0.001 19 SRKLKRIKK 0.001 30 EKSQFLEEG 0.001 22 LKRIKKGWE 0.001 25 IKKGWEKSQ 0.001 36 EEGIGGTIP 0.001

TABLE XVI-V7A HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 SPKSLSETF 2.400 9 FLPNGINGI 1.800 4 SLSETFLPN 0.144 6 SETFLPNGI 0.144 7 ETFLPNGIN 0.100 8 TFLPNGING 0.090 2 PKSLSETFL 0.040 3 KSLSETFLP 0.030 5 LSETFLPNG 0.015

TABLE XVI-V7B HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 5 AYQQSTLGY 7.500 9 STLGYVALL 6.000 8 QSTLGYVAL 4.000 3 NMAYQQSTL 4.000 1 FLNMAYQQS 0.180 2 LNMAYQQST 0.180 6 YQQSTLGYV 0.150 7 QQSTLGYVA 0.120 4 MAYQQSTLG 0.010

TABLE XVI-V7C HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 139 KSQAASGTL 12.000 29 RGGLSEIVL 8.000 181 KSKHCMFSL 8.000 130 LWEFLLRLL 7.200 24 GANILRGGL 7.200 127 VGPLWEFLL 6.000 126 GVGPLWEFL 5.760 152 TSWSLGEFL 4.800 160 LGSGTWMKL 4.400 148 SLAFTSWSL 4.000 42 QQDRKIPPL 4.000 15 SPAAAWKCL 4.000 141 QAASGTLSL 4.000 5 ILDLSVEVL 4.000 165 WMKLETIIL 4.000 113 ANSWRNPVL 4.000 158 EFLGSGTWM 3.750 125 NGVGPLWEF 3.300 143 ASGTLSLAF 2.400 151 FTSWSLGEF 2.200 179 EQKSKHCMF 2.000 164 TWMKLETII 1.800 31 GLSEIVLPI 1.680 66 TAEAQESGI 1.500 19 AWKCLGANI 1.200 27 ILRGGLSEI 1.100 163 GTWMKLETI 1.000 132 EFLLRLLKS 0.825 168 LETIILSKL 0.616 102 PPESPDRAL 0.600 50 LSTPPPPAM 0.600 129 PLWEFLLRL 0.480 20 WKCLGANIL 0.480 108 RALKAANSW 0.360 117 RNPVLPHTN 0.360 136 RLLKSQAAS 0.300 82 SQIPVVGVV 0.252 4 VILDLSVEV 0.238 123 HTNGVGPLW 0.210 83 QIPVVGVVT 0.210 104 ESPDRALKA 0.198 51 STPPPPAMW 0.180 145 GTLSLAFTS 0.180 154 WSLGEFLGS 0.180 68 EAGESGIRN 0.180 9 SVEVLASPA 0.180 59 WTEEAGATA 0.180 156 LEGFLGSGT 0.180 52 TPPPPAMWT 0.180 112 AANSWRNPV 0.180 101 DPPESPDRA 0.180 2 SIVILDLSV 0.180 169 ETIILSKLT 0.180 88 GVVTEDDEA 0.165 14 ASPAAAWKC 0.165 25 ANILRGGLS 0.150 72 SGIRNKSSS 0.150 11 EVLASPAAA 0.150 81 SSQIPVVGV 0.150 177 TQEQKSKHC 0.150 147 LSLAFTSWS 0.150 64 GATAEAQES 0.132 134 LLRLLKSQA 0.120 146 TLSLAFTSW 0.120 185 CMFSLISGS 0.120 182 SKHCMFSLI 0.120 58 MWTEEAGAT 0.120 92 EDDEAQDSI 0.120 39 IEWQQDRKI 0.110 162 SGTWMKLET 0.110 17 AAAWKCLGA 0.100 79 SSSSQIPVV 0.100 140 SQAASGTLS 0.100 76 NKSSSSSQI 0.100 142 AASGTLSLA 0.100 105 SPDRALKAA 0.100 57 AMWTEEAGA 0.100 144 SGTLSLAFT 0.100 18 AAWKCLGAN 0.100 7 DLSVEVLAS 0.100 78 SSSSSQIPV 0.100 12 VIASPAAAW 0.100 73 GIRNKSSSS 0.100 71 ESGIRNKSS 0.100 178 QEQKSKHCM 0.075 150 AFTSWSLGE 0.050 46 KIPPLSTPP 0.043 167 KLETIILSK 0.042 122 PHTNGVGPL 0.040 21 KCLGANILR 0.030 116 WRNPVLPHT 0.025 35 IVLPIEWQQ 0.025 8 LSVEVLASP 0.025 77 KSSSSSQIP 0.024 119 PVLPHTNGV 0.022 37 LPIEWQQDR 0.022 1 PSIVILDLS 0.021 6 LDLSVEVLA 0.021 32 LSEIVLPIE 0.021 183 KHCMFSLIS 0.020

TABLE XVII-V1 HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 124 QYPESNAEYL 360.000 359 MYISFGIMSL 300.000 399 GYVALLISTF 180.000 282 LYYGTKYRRF 100.000 423 YYRFYTPPNF 100.000 290 RFPPWLETWL 86.400 425 RFYTPPNFVL 40.000 186 NFIPIDLGSL 36.000 145 GFNVVSAWAL 30.000 40 DFAKSLTIRL 24.000 257 KTLPIVAITL 20.160 362 SFGIMSLGLL 20.000 213 RGPVVVAISL 16.800 183 RQLNFIPIDL 16.800 377 TSIPSVSNAL 12.096 131 EYLASLFPDS 10.800 250 IPIEIVNKTL 10.080 238 PYARNQQSDF 10.000 270 VYLAGLLAAA 9.000 437 VLPSIVILDL 8.400 312 FAMVHVAYSL 8.400 279 AYQLYYGTKY 8.250 165 VYICSNNIQA 7.500 176 QQVIELARQL 7.200 202 ENLPLRLFTL 7.200 99 TSLWDLRHLL 7.200 427 YTPPNFVLAL 7.200 303 KQLGLLSFFF 7.200 267 LSLVYLAGLL 7.200 426 FYTPPNFVLA 7.200 402 ALLISTFHVL 7.200 53 GYHVVIGSRN 7.000 247 FYKIPIEIVN 7.000 364 GIMSLGLLSL 6.000 127 ESNAEYLASL 6.000 61 RNPKFASEFF 6.000 298 WLQCRKQLGL 6.000 4 ISMMGSPKSL 6.000 273 AGLLAAAYQL 6.000 323 LPMRRSERYL 6.000 147 NVVSAWALQL 6.000 435 ALVLPSIVIL 6.000 440 SIVILDLLQL 6.000 258 TLPIVAITLL 6.000 438 LPSIVILDLL 5.600 422 EYYRFYTPPN 5.000 219 AISLATFFFL 4.800 417 RAFEEEYYRF 4.800 365 IMSLGLLSLL 4.800 197 SAREIENLPL 4.800 172 IQARQQVIEL 4.400 356 RIEMYISFGI 4.200 36 IGSSDFAKSL 4.000 98 YTSLWDLRHL 4.000 132 YLASLFPDSL 4.000 296 ETWLQCRKQL 4.000 266 LLSLVYLAGL 4.000 195 LSSAREIENL 4.000 314 MVHVAYSLCL 4.000 263 AITLLSLVYL 4.000 299 LQCRKQLGLL 4.000 92 AIHREHYTSL 4.000 361 ISFGIMSLGL 4.000 9 SPKSLSETCL 4.000 395 QSTLGYVALL 4.000 394 IQSTLGYVAL 4.000 241 RNQQSDFYKI 3.960 163 RQVYICSNNI 3.600 382 VSNALNWREF 3.300 56 VVIGSRNPKF 3.300 384 NALNWREFSF 3.000 410 VLIYGWKRAF 3.000 216 VVVAISLATF 3.000 178 VIELARQLNF 3.000 218 VAISLATFFF 3.000 200 EIENLPLRLF 3.000 81 DALTKTNIIF 3.000 128 SNAEYLASLF 2.880 137 FPDSLIVKGF 2.800 111 KILIDVSNNM 2.520 217 VVAISLATFF 2.400 16 TCLPNGINGI 2.160 327 RSERYLFLNM 2.160 13 LSETCLPNGI 2.160 396 STLGYVALLI 2.100 432 FVLALVLPSI 2.100 354 VWRIEMYISF 2.000 222 LATFFFLYSF 2.000 32 TVGVIGSGDF 2.000 385 ALNWREFSFI 1.800 170 NNIQARQQVI 1.800 348 SWNEEEVWRI 1.800 199 REIENLPLRL 1.728 403 LLISTFHVLI 1.500 330 RYLFLNMAYQ 1.500 434 LALVLPSIVI 1.500 211 LWRGPVVVAI 1.400 336 MAYQQVHANI 1.400 227 FLYSFVRDVI 1.400 103 DLRHLLVGKI 1.320

TABLE XVII-V2 HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 16 GFTPFSCLSL 24.000 32 RCPPPCPADF 7.200 2 GSPGLQALSL 6.000 30 DYRCPPPCPA 5.000 14 SSGFTPFSCL 4.800 11 LSLSSGFTPF 3.600 33 CPPPCPADFF 3.600 8 ALSLSLSSGF 2.400 4 PGLQALSLSL 0.720 34 PPPCPADFFL 0.600 36 PCPADFFLYF 0.360 9 LSLSLSSFGT 0.150 5 GLQALSLSLS 0.150 24 SLPSSWDYRC 0.150 1 SGSPGLWALS 0.144 6 LQALSLSLSS 0.120 20 FSCLSLPSSW 0.120 18 TFPSCLSLPS 0.120 15 SGFTPFSCLS 0.100 12 SLSSGFTPFS 0.100 28 SWDYRCPPPC 0.100 13 LSSGFTPFSC 0.100 3 SPGLQALSLS 0.100 22 CLSLPSSWDY 0.100 19 PFSCLSLPSS 0.050 23 LSLPSSWDYR 0.018 7 QALSLSLSSG 0.015 17 FTPFSCLSLP 0.015 21 SCLSLPSSWD 0.015 35 PPCPADFFLY 0.014 27 SSWDYRCPPP 0.012 25 LPSSWDYRCP 0.010 10 SLSLSSGFTP 0.010 31 YRCPPPCPAD 0.001 29 WDYRCPPPCP 0.001 26 PSSWDYRCPP 0.001

TABLE XVII-V5A HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 1 ENLPLRLFTF 3.600 10 FWRGPVVVAI 1.400 7 LFTFWRGPVV 0.500 9 TFWRGPVVVA 0.500 2 NLPLRLFTFW 0.216 6 RLFTFWRGPV 0.200 8 FTFWRGPVVV 0.100 3 LPLRLTFTWR 0.015 5 LRLTFTWRGP 0.002 4 PLRLFTFWRG 0.001

TABLE XVII-V5B HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 24 EFVFLLTLLL 36.000 22 ELEFVFLLTL 6.000 20 ELELEFVFLL 6.000 12 CSFADTQTEL 4.400 14 FADTQTELEL 4.400 16 DTQTELELEF 3.960 18 QTELELEFVF 3.600 5 FSFIQIFCSF 3.360 1 NWREFSFIQI 1.440 19 TELELEFVFL 0.864 6 SFIQIFCSFA 0.750 4 EFSFIQIFCS 0.500 10 IFCSFADTQT 0.500 23 LEFVFLLTLL 0.480 2 WREFSFIQIF 0.360 8 IQIFCSFADT 0.180 17 TQTELELEFV 0.120 13 SFADTQTELE 0.060 21 LELEFVFLLT 0.030 3 REFSFIQIFC 0.028 7 FIQIFCSFAD 0.015 11 FCSFADTQTE 0.012 9 QIFCSFADTQ 0.010 15 ADTQTELELE 0.001

TABLE XVII-V6 HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 14 LFLPCISRKL 55.440 7 VILGKIILFL 8.400 5 SIVILGKIIL 6.000 6 IVILGKIILF 3.000 35 FLEEGIGGTI 2.520 3 LPSIVILGKI 1.540 27 KKGWEKSQFL 0.960 34 QFLEEGIGGT 0.900 46 HVSPERVTVM 0.600 11 KIILFLPCIS 0.360 26 IKKGWEKSQF 0.200 4 PSIVILGKII 0.180 38 EGIGGTIPHV 0.150 17 PCISRKLKRI 0.150 43 TIPHVSPERV 0.150 10 GKIILFLPCI 0.150 9 LGKIILFLPC 0.144 39 GIGGTIPHVS 0.140 31 EKSQFLEEGI 0.120 44 IPHVSPERVT 0.100 21 RKLKRIKKGW 0.042 24 KRIKKGWEKS 0.033 32 KSQFLEEGIG 0.030 42 GTIPHVSPER 0.028 1 LVLPSIVILG 0.025 28 KGWEKSQFLE 0.024 2 VLPSIVILGK 0.021 25 RIKKGWEKSQ 0.020 22 KLKRIKKGWE 0.020 29 GWEKSQFLEE 0.020 12 IILFLPCISR 0.015 15 FLPCISRKLK 0.015 8 ILGKIILFLP 0.014 18 CISRKLKRIK 0.012 16 LPCISRKLKR 0.011 19 ISRKLKRIKK 0.011 33 SQFLEEGIGG 0.010 41 GGTIPHVSPE 0.010 40 IGGTIPHVSP 0.010 13 ILFLPCISRK 0.010 36 LEEGIGGTIP 0.002 45 PHVSPERVTV 0.002 20 SRKLKRIKKG 0.001 30 WEKSQFLEEG 0.001 23 LKRIKKGWEK 0.001 37 EEGIGGTIPH 0.001

TABLE XVII-V7A HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 9 TFLPNGINGI 10.800 2 SPKSLSETFL 4.000 1 GSPKSLSETF 3.600 6 LSETFLPNGI 2.160 4 KSLSETFLPN 0.360 10 FLPNGINGIK 0.021 5 SLSETFLPNG 0.012 7 SETFLPNGIN 0.010 8 ETFLPNGING 0.010 3 PKSLSETFLP 0.000

TABLE XVII-V7B A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 6 AYQQSTLGYV 7.500 3 LNMAYQQSTL 6.000 8 QQSTLGYVAL 4.000 9 QSTLGYVALL 4.000 10 STLGYVALLI 2.100 1 LFLNMAYQQS 0.900 7 YQQSTLGYVA 0.180 2 FLNMAYQQST 0.180 5 MAYQQSTLGY 0.100 4 NMAYQQSTLG 0.010

TABLE XVIl-V7C HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 168 KLETIILSKL 18.480 151 AFTSWSLGEF 11.000 5 VILDLSVEVL 7.200 42 WQQDRKIPPL 7.200 126 NGVGPLWEFL 7.200 102 DPPESPDRAL 7.200 113 AANSWRNPVL 6.000 129 GPLWEFLLRL 6.000 148 LSLAFTSWSL 6.000 15 ASPAAAWKCL 6.000 165 TWMKLETIIL 6.000 24 LGANILRGGL 4.800 20 AWKCLGANIL 4.800 127 GVGPLWEFLL 4.800 152 FTSWSLGEFL 4.800 160 FLGSGTWMKL 4.400 122 LPHTNGVGPL 4.000 141 SQAASGTLSL 4.000 182 KSKHCMFSLI 2.400 143 AASGTLSLAF 2.400 125 TNGVGPLWEF 2.200 31 GGLSEIVLPI 2.100 76 RNKSSSSSQI 2.000 27 NILRGGLSEI 1.650 164 GTWMKLETII 1.200 19 AAWKCLGANI 1.200 66 ATAEAQESGI 1.200 163 SGTWMKLETI 1.000 178 TQEQKSKHCM 0.750 130 PLWEFLLRLL 0.576 29 LRGGLSEIVL 0.400 181 QKSKHCMFSL 0.400 139 LKSQAASGTL 0.400 179 QEQKSKHCMF 0.300 140 KSQAASGTLS 0.300 70 AQESGIRNKS 0.277 83 SQIPVVGVVT 0.252 112 KAANSWRNPV 0.240 91 VTEDDEAQDS 0.216 9 LSVEVLASPA 0.216 82 SSQIPVVGVV 0.210 78 KSSSSSQIPV 0.200 4 IVILDLSVEV 0.198 33 LSEIVLPIEW 0.198 119 NPVLPHTNGV 0.180 105 ESPDRALKAA 0.180 52 STPPPPAMWT 0.180 177 LTQEQKSKHC 0.180 134 FLLRLLKSQA 0.180 185 HCMFSLISGS 0.180 146 GTLSLAFTSW 0.180 39 PIEWQQDRKI 0.165 88 VGVVTEDDEA 0.165 10 SVEVLASPAA 0.150 73 SGIRNKSSSS 0.150 25 GANILRGGLS 0.150 157 LGEFLGSGTW 0.150 12 EVLASPAAAW 0.150 156 SLGEFLGSGT 0.144 1 LPSIVILDLS 0.140 6 ILDLSVEVLA 0.140 116 SWRNPVLPHT 0.140 43 QQDRKIPPLS 0.140 64 AGATAEAQES 0.132 14 LASPAAAWKC 0.132 174 LSKLTQEQKS 0.132 51 LSTPPPPAMW 0.120 92 TEDDEAQDSI 0.120 135 LLRLLKSQAA 0.120 106 SPDRALKAAN 0.120 59 MWTEEAGATA 0.120 28 ILRGGLSEIV 0.120 154 SWSLGEFLGS 0.120 145 SGTLSLAFTS 0.120 162 GSGTWMKLET 0.110 97 AQDSIDPPES 0.110 147 TLSLAFTSWS 0.100 180 EQKSKHCMFS 0.100 79 SSSSSQIPVV 0.100 142 QAASGTLSLA 0.100 18 AAAWKCLGAN 0.100 138 LLKSQAASGT 0.100 110 ALKAANSWRN 0.100 144 ASGTLSLAFT 0.100 74 GIRNKSSSSS 0.100 81 SSSQIPVVGV 0.100 166 WMKLETIILS 0.100 72 ESGIRNKSSS 0.100 58 AMWTEEAGAT 0.100 133 EFLLRLLKSQ 0.090 159 EFLGSGTWMK 0.075 158 GEFLGSGTWM 0.050 50 PLSTPPPPAM 0.050 47 KIPPLSTPPP 0.036 22 KCLGANILRG 0.030 118 RNPVLPHTNG 0.030 109 RALKAANSWR 0.030 137 RLLKSQAASG 0.030 96 EAGDSIDPPE 0.025 172 IILSKLTQEQ 0.024

TABLE XVIII-V1 HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 173 QARQQVIEL 120.000 214 GPVVVAISL 80.000 259 LPIVAITLL 80.000 428 TPPNFVLAL 80.000 438 LPSIVILDL 80.000 291 FPPWLETWL 80.000 300 QCRKQLGLL 40.000 125 YPESNAEYL 24.000 177 QVIELARQL 20.000 148 VVSAWALQL 20.000 261 IVAITLLSL 20.000 75 DVTHHEDAL 20.000 441 IVILDLLQL 20.000 436 LVLPSIVIL 20.000 41 FAKSLTIRL 12.000 313 AMVHVAYSL 12.000 133 LASLFPDSL 12.000 5 SMMGSPKSL 12.000 27 DARKVTVGV 6.000 100 SLWDLRHLL 6.000 146 FNVVSAWAL 4.000 220 ISLATFFFL 4.000 187 FIPIDLGSL 4.000 128 SNAEYLASL 4.000 363 FGIMSLGLL 4.000 274 GLLAAAYQL 4.000 365 IMSLGLLSL 4.000 366 MSLGLLSLL 4.000 184 QLNFIPIDL 4.000 93 IHREHYTSL 4.000 324 PMRRSERYL 4.000 395 QSTLGYVAL 4.000 267 LSLVYLAGL 4.000 268 SLVYLAGLL 4.000 360 YISFGIMSL 4.000 196 SSAREIENL 4.000 378 SIPSVSNAL 4.000 258 TLPIVAITL 4.000 299 LQCRKQLGL 4.000 99 TSLWDLRHL 4.000 403 LLISTFHVL 4.000 37 GSGDFAKSL 4.000 203 NLPLRLFTL 4.000 264 ITLLSLVYL 4.000 396 STLGYVALL 4.000 287 KYRRFPPWL 4.000 157 GPKDASRQV 4.000 317 VAYSLCLPM 3.000 9 SPKSLSETC 2.000 250 IPIEIVNKT 2.000 353 EVWRIEMYI 2.000 49 LIRCGYHVV 2.000 164 QVYICSNNI 2.000 134 ASLFPDSLI 1.800 435 ALVLPSIVI 1.800 200 EIENLPLRL 1.200 81 DALTKTNII 1.200 323 LPMRRSERY 1.200 108 LVGKILIDV 1.000 358 EMYISFGIM 1.000 112 ILIDVSNNM 1.000 254 IVNKTLPIV 1.000 231 FVRDVIHPY 1.000 328 SERYLFLNM 1.000 306 GLLSFFFAM 1.000 278 AAYQLYYGT 0.900 402 ALLISTFHV 0.600 297 TWLQCRKQL 0.600 262 VAITLLSLV 0.600 239 YARNQQSDF 0.600 434 LALVLPSIV 0.600 65 FASEFFPHV 0.600 161 ASRQVYICS 0.600 426 FYTPPNFVL 0.600 374 LAVTSIPSV 0.600 314 MVHVAYSLC 0.500 34 GVIGSGDFA 0.500 216 VVVAISLAT 0.500 269 LVYLAGLLA 0.500 237 HPYARNQQS 0.400 371 LSLLAVTSI 0.400 85 KTNIIFVAI 0.400 390 EFSFIQSTL 0.400 439 PSIVILDLL 0.400 397 TLGYVALLI 0.400 430 PNFVLALVL 0.400 362 SFGIMSLGL 0.400 171 NIQARQQVI 0.400 180 ELARQLNFI 0.400 193 GSLSSAREI 0.400 386 LNWREFSFI 0.400 204 LPLRLFTLW 0.400 429 PPNFVLALV 0.400 188 IPIDLGSLS 0.400 379 IPSVSNALN 0.400 62 NPKFASEFF 0.400 326 RRSERYLFL 0.400 433 VLALVLPSI 0.400 253 EIVNKTLPI 0.400 106 HLLVGKILI 0.400

TABLE XVIlI-V2 HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 3 SPGLQALSL 80.000 35 PPCPADFFL 8.000 15 SGFTPFSCL 6.000 1 SGSPGLQAL 4.000 17 FTPFSCLSL 4.000 5 GLQALSLSL 4.000 25 LPSSWDYRC 2.000 37 CPADFFLYF 0.400 33 CPPPCPADF 0.400 18 TPFSCLSLP 0.200 10 SLSLSSGFT 0.100 14 SSGFTPFSC 0.100 7 QALSLSLSS 0.060 34 PPPCPADFF 0.060 8 ALSLSLSSG 0.030 23 LSLPSSWDY 0.020 12 SLSSGFTPF 0.020 21 SCLSLPSSW 0.020 6 LQALSLSLS 0.020 13 LSSGFTPFS 0.020 2 GSPGLQALS 0.020 9 LSLSLSSGF 0.020 20 FSCLSLPSS 0.020 32 RCPPPCPAD 0.015 22 CLSLPSSWD 0.015 31 YRCPPPCPA 0.015 30 DYRCPPPCP 0.015 27 SSWDYRCPP 0.015 29 WDYRCPPPC 0.010 24 SLPSSWDYR 0.010 11 LSLSSGFTP 0.010 36 PCPADFFLY 0.002 16 GFTPFSCLS 0.002 4 PGLQALSLS 0.002 26 PSSWDYRCP 0.001 28 SWDYRCPPP 0.000 19 PFSCLSLPS 0.000

TABLE XVIII-V5A HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 2 LPLRLFTFW 0.400 7 FTFWRGPVV 0.200 9 FWRGPVVVA 0.150 6 LFTFWRGPV 0.030 8 TFWRGPVVV 0.020 1 NLPLRLFTF 0.020 3 PLRLFTFWR 0.010 5 RLFTFWRGP 0.010 4 LRLFTFWRG 0.001

TABLE XVIII-V5B HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 24 FVFLLTLLL 20.000 14 ADTQTELEL 1.200 19 ELELEFVFL 1.200 12 SFADTQTEL 0.400 23 EFVFLLTLL 0.400 22 LEFVFLLTL 0.400 20 LELEFVFLL 0.400 10 FCSFADTQT 0.100 8 QIFCSFADT 0.100 6 FIQIFCSFA 0.100 17 QTELELEFV 0.060 21 ELEFVFLLT 0.030 4 FSFIQIFCS 0.020 16 TQTELELEF 0.020 1 WREFSFIQI 0.012 11 CSFADTQTE 0.010 3 EFSFIQIFC 0.010 7 IQIFCSFAD 0.010 15 DTQTELELE 0.010 13 FADTQTELE 0.009 5 SFIQIFCSF 0.002 2 REESFIQIF 0.002 18 TELELEFVF 0.002 9 IFCSFADTQ 0.001

TABLE XVIII-V6 HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 5 IVILGKIIL 20.000 14 FLPCISRKL 4.000 43 IPHVSPERV 4.000 7 ILGKIILFL 4.000 27 KGWEKSQFL 4.000 45 HVSPERVTV 1.500 46 VSPERVTVM 1.000 31 KSQFLEEGI 0.400 4 SIVILGKII 0.400 17 CISRKLKRI 0.400 10 KIILFLPCI 0.400 15 LPCISRKLK 0.300 38 GIGGTIPHV 0.200 2 LPSIVILGK 0.200 18 ISRKLKRIK 0.100 3 PSIVILGKI 0.040 34 FLEEGIGGT 0.030 11 IILFLPCIS 0.020 39 IGGTIPHVS 0.020 6 VILGKIILF 0.020 24 RIKKGWEKS 0.020 21 KLKRIKKGW 0.020 40 GGTIPHVSP 0.015 12 ILFLPCISR 0.015 35 LEEGIGGTI 0.012 37 EGIGGTIPH 0.010 22 LKRIKKGWE 0.010 8 LGKIILFLP 0.010 32 SQELEEGIG 0.010 41 GTIPHVSPE 0.010 1 VLPSIVILG 0.010 9 GKIILFLPC 0.010 42 TIPHVSPER 0.010 26 KKGWEKSQF 0.002 19 SRKLKRIKK 0.002 44 PHVSPERVT 0.002 36 EEGIGGTIP 0.001 20 RKLKRIKKG 0.001 29 WEKSQFLEE 0.001 13 LFLPCISRK 0.001 25 IKKGWEKSQ 0.001 30 EKSQFLEEG 0.001

TABLE XVlII-V7A HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 FLPNGINGI 0.400 1 SPKSLSETF 0.400 6 SETFLPNGI 0.040 2 PKSLSETFL 0.040 7 ETFLPNGIN 0.030 4 SLSETFLPN 0.020 3 KSLSETFLP 0.010 5 LSETFLPNG 0.003 8 TFLPNGING 0.001

TABLE XVIII-V7B HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 STLGYVALL 4.000 8 QSTLGYVAL 4.000 3 NMAYQQSTL 4.000 2 LNMAYQQST 0.300 6 YQQSTLGYV 0.200 7 QQSTLGYVA 0.100 4 MAYQQSTLG 0.030 1 FLNMAYQQS 0.020 5 AYQQSTLGY 0.006

TABLE XVIII-V7C HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 15 SPAAAWKCL 80.000 126 GVGPLWEFL 20.000 24 GANILRGGL 18.000 113 ANSWRNPVL 12.000 141 QAASGTLSL 12.000 127 VGPLWEFLL 4.000 148 SLAFTSWSL 4.000 181 KSKHCMFSL 4.000 29 RGGLSEIVL 4.000 139 KSQAASGTL 4.000 27 ILRGGLSEI 4.000 165 WMKLETIIL 4.000 152 TSWSLGEFL 4.000 160 LGSGTWMKL 4.000 102 PPESPDRAL 3.600 52 TPPPPAMWT 3.000 112 AANSWRNPV 2.700 101 DPPESPDRA 2.000 50 LSTPPPPAM 1.500 5 ILDLSVEVL 1.200 42 QQDRKIPPL 1.200 134 LLRLLKSQA 1.000 142 AASGTLSLA 0.900 17 AAAWKCLGA 0.900 105 SPDRALKAA 0.600 11 EVLASPAAA 0.500 88 GVVTEDDEA 0.500 31 GLSEIVLPI 0.400 20 WKCLGANIL 0.400 168 LETIILSKL 0.400 163 GTWMKLETI 0.400 129 PLWEFLLRL 0.400 66 TAEAQESGI 0.360 81 SSQIPVVGV 0.300 57 AMWTEEAGA 0.300 14 ASPAAAWKC 0.300 118 NPVLPHTNG 0.300 84 IPVVGVVTE 0.200 79 SSSSQIPVV 0.200 55 PPAMWTEEA 0.200 82 SQIPVVGVV 0.200 37 LPIEWQQDR 0.200 78 SSSSSQIPV 0.200 73 GIRNKSSSS 0.200 4 VILDLSVEV 0.200 2 SIVILDLSV 0.200 47 IPPLSTPPP 0.200 128 GPLWEFLLR 0.200 121 LPHTNGVGP 0.200 18 AAWKCLGAN 0.180 9 SVEVLASPA 0.150 164 TWMKLETII 0.120 19 AWKCLGANI 0.120 130 LWEFLLRLL 0.120 104 ESPDRALKA 0.100 158 EFLGSGTWM 0.100 162 SGTWMKLET 0.100 169 ETIILSKLT 0.100 83 QIPVVGVVT 0.100 178 QEQKSKHCM 0.100 144 SGTLSLAFT 0.100 119 PVLPHTNGV 0.100 143 ASGTLSLAF 0.060 64 GATAEAQES 0.060 68 EAQESGIRN 0.060 25 ANILRGGLS 0.060 108 RALKAANSW 0.060 35 IVLPIEWQQ 0.050 86 VVGVVTEDD 0.050 3 IVILDLSVE 0.050 89 VVTEDDEAQ 0.050 122 PHTNGVGPL 0.040 76 NKSSSSSQI 0.040 182 SKHCMFSLI 0.040 39 IEWQQDRKI 0.040 12 VLASPAAAW 0.030 62 EAGATAEAQ 0.030 125 NGVGPLWEF 0.030 13 LASPAAAWK 0.030 109 ALKAANSWR 0.030 63 AGATAEAQE 0.030 95 EAQDSIDPP 0.030 65 ATAEAQESG 0.030 149 LAFTSWSLG 0.030 111 KAANSWRNP 0.030 51 STPPPPAMW 0.030 184 HCMFSLISG 0.030 59 WTEEAGATA 0.030 156 LGEFLGSGT 0.030 177 TQEQKSKHC 0.030 140 SQAASGTLS 0.020 48 PPLSTPPPP 0.020 71 ESGIRNKSS 0.020 123 HTNGVGPLW 0.020 72 SGIRNKSSS 0.020 179 EQKSKHCMF 0.020 185 CMFSLISGS 0.020 54 PPPAMWTEE 0.020 147 LSLAFTSWS 0.020 28 LRGGLSEIV 0.020

TABLE XIX-V1 HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 332 LPMRRSERYL 240.000 197 SAREIENLPL 120.000 438 LPSIVILDLL 80.000 9 SPKSLSETCL 80.000 250 IPIEIVNKTL 80.000 312 FAMVHVAYSL 36.000 147 NVVSAWALQL 20.000 314 MVHVAYSLCL 20.000 364 GIMSLGLLSL 12.000 263 AITLLSLVYL 12.000 219 AISLATFFFL 12.000 402 ALLISTFHVL 12.000 435 ALVLPSIVIL 12.000 273 AGLLAAAYQL 12.000 4 ISMMGSPKSL 12.000 92 AIHREHYTSL 12.000 27 DARKVTVGVI 12.000 181 LARQLNFIPI 12.000 429 PPNFVLALVL 8.000 296 ETWLQCRKQL 6.000 99 TSLWDLRHLL 6.000 316 HVAYSLCLPM 5.000 231 FVRDVIHPYA 5.000 195 LSSAREIENL 4.000 257 KTLPIVAITL 4.000 377 TSIPSVSNAL 4.000 266 LLSLVYLAGL 4.000 202 ENLPLRLFTL 4.000 132 YLASLFPDSL 4.000 299 LQCRKQLGLL 4.000 176 QQVIELARQL 4.000 427 YTPPNFVLAL 4.000 394 IQSTLGYVAL 4.000 213 RGPVVVAISL 4.000 365 IMSLGLLSLL 4.000 49 LIRCGYHVVI 4.000 428 TPPNFVLALV 4.000 103 DLRHLLVGKI 4.000 36 IGSGDFAKSL 4.000 98 YTSLWDLRHL 4.000 298 WLQCRKQLGL 4.000 325 MRRSERYLFL 4.000 361 ISFGIMSLGL 4.000 258 TLPIVAITLL 4.000 172 IQARQQVIEL 4.000 127 ESNAEYLASL 4.000 440 SIVILDLLQL 4.000 183 RQLNFIPIDL 4.000 267 LSLVYLAGLL 4.000 437 VLPSIVILDL 4.000 395 QSTLGYVALL 4.000 173 QARQQVIELA 3.000 432 FVLALVLPSI 2.000 214 GPVVVAISLA 2.000 434 LALVLPSIVI 1.800 133 LASLFPDSLI 1.800 385 ALNWREFSFI 1.200 336 MAYQQVHANI 1.200 41 FAKSLTIRLI 1.200 111 KILIDVSNNM 1.000 261 IVAITLLSLV 1.000 305 LGLLSFFFAM 1.000 277 AAAYQLYYGT 0.900 161 ASRQVYICSN 0.600 239 YARNQQSDFY 0.600 255 VNKTLPIVAI 0.600 401 VALLISTFHV 0.600 125 YPESNAEYLA 0.600 157 GPKDASRQVY 0.600 227 FLYSFVRDVI 0.600 82 ALTKTNIIFV 0.600 425 RFYTPPNFVL 0.600 65 FASEFFPHVV 0.600 134 ASLFPDSLIV 0.600 223 ATFFFLYSFV 0.600 269 LVYLAGLLAA 0.500 142 IVKGFNVVSA 0.500 75 DVTHHEDALT 0.500 441 IVILDLLQLC 0.500 409 HVLIYGWKRA 0.500 254 IVNKTLPIVA 0.500 90 FVAIHREHYT 0.500 375 AVTSIPSVSN 0.450 199 REIENLPLRL 0.400 95 REHYTSLWDL 0.400 379 IPSVSNALNW 0.400 259 LPIVAITLLS 0.400 211 LWRGPVVVAI 0.400 163 RQVYICSNNI 0.400 145 GFNVVSAWAL 0.400 186 NFIPIDLGSL 0.400 188 IPIDLGSLSS 0.400 370 LLSLLAVTSI 0.400 359 MYISFGIMSL 0.400 16 TCLPNGINGI 0.400 124 QYPESNAEYL 0.400 170 NNIQARQQVI 0.400 243 QQSDFYKIPI 0.400 241 RNQQSDFYKI 0.400 74 VDVTHHEDAL 0.400

TABLE XIX-V2 HLA-B7-10mers-98P486 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 34 PPPCPADFFL 8.000 14 SSGFTPFSCL 6.000 2 GSPGLQALSL 4.000 33 CPPPCPADFF 0.600 18 TPFSCLSLPS 0.400 16 GFTPFSCLSL 0.400 3 SPGLQALSLS 0.400 4 PGLQALSLSL 0.400 25 LPSSWDYRCP 0.200 30 DYRCPPPCPA 0.150 24 SLPSSWDYRC 0.100 13 LSSGFTPFSC 0.100 9 LSLSLSSGFT 0.100 8 ALSLSLSSGF 0.060 35 PPCPADFFLY 0.040 7 QALSLSLSSG 0.030 15 SGFTPFSCLS 0.020 22 CLSLPSSWDY 0.020 11 LSLSSGFTPF 0.020 6 LQALSLSLSS 0.020 32 RCPPPCPADF 0.020 1 SGSPGLQALS 0.020 20 FSCLSLPSSW 0.020 12 SLSSGFTPFS 0.020 5 GLQALSLSLS 0.020 21 SCLSLPSSWD 0.015 10 SLSLSSGFTP 0.010 17 FTPFSCLSLP 0.010 27 SSWDYRCPPP 0.010 23 LSLPSSWDYR 0.010 28 SWDYRCPPPC 0.003 36 PCPADEFLYF 0.002 26 PSSWDYRCPP 0.002 31 YRCPPPCPAD 0.002 29 WDYRCPPPCP 0.002 19 PFSCLSLPSS 0.000

TABLE XIX-V5A HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 10 FWRGPVVVAI 0.400 6 RLFTFWRGPV 0.300 8 FTFWRGPVVV 0.200 3 LPLRLFTFWR 0.200 2 NLPLRLFTFW 0.020 7 LFTFWRGPVV 0.020 1 ENLPLRLFTF 0.020 9 TFWRGPVVVA 0.015 4 PLRLFTFWRG 0.010 5 LRLFTFWREGP 0.001

TABLE XIX-V5B HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 12 CSFADTQTEL 4.000 14 FADTQTELEL 3.600 20 ELELEFVFLL 1.200 22 ELEFVFLLTL 1.200 23 LEFVFLLTLL 0.400 1 NWREFSFIQI 0.400 19 TELELEFVFL 0.400 24 EFVFLLTLLL 0.400 17 TQTELELEFV 0.200 8 IQIFCSFADT 0.100 5 FSFIQIFCSF 0.020 16 DTQTELELEF 0.020 10 IFCSFADTQT 0.010 21 LELEFVFLLT 0.010 6 SFIQIFCSFA 0.010 3 REFSFIQIFC 0.010 9 QIFCSFADTQ 0.010 7 FIQIFCSFAD 0.010 11 FCSFADTQTE 0.010 18 QTELELEFVF 0.006 15 ADTQTELELE 0.003 4 EFSFIQIFCS 0.002 13 SFADTQTELE 0.001 2 WREFSFIQIF 0.001

TABLE XIX-V6 HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 3 LPSIVILGKI 8.000 46 HVSPERVTVM 5.000 5 SIVILGKIIL 4.000 7 VILGKIILFL 4.000 44 IPHVSPERVT 3.000 14 LFLPCISRKL 0.400 27 KKGWEKSQFL 0.400 16 LPCISRKLKR 0.200 43 TIPHVSPERV 0.200 38 EGIGGTIPHV 0.200 19 ISRKLKRIKK 0.150 35 FLEEGIGGTI 0.120 9 LGKIILFLPC 0.100 6 IVILGKIILF 0.100 1 LVLPSIVILG 0.050 10 GKIILFLPCI 0.040 4 PSIVILGKII 0.040 31 EKSQFLEEGI 0.040 17 PCISRKLKRI 0.040 11 KIILFLPCIS 0.020 39 GIGGTIPHVS 0.020 15 FLPCISRKLK 0.015 40 IGGTIPHVSP 0.015 12 IILFLPCISR 0.015 34 QFLEEGIGGT 0.010 2 VLPSIVILGK 0.010 33 SQFLEEGIGG 0.010 25 RIKKGWEKSQ 0.010 32 KSQFLEEGIG 0.010 13 ILFLPCISRK 0.010 22 KLKRIKKGWE 0.010 8 ILGKIILFLP 0.010 41 GGTIPHVSPE 0.010 18 CISRKLKRIK 0.010 28 KGWEKSQFLE 0.010 42 GTIPHVSPER 0.010 23 LKRIKKGWEK 0.010 45 PHVSPERVTV 0.003 24 KRIKKGWEKS 0.002 26 IKKGWEKSQF 0.002 21 RKLKRIKKGW 0.002 20 SRKLKRIKKG 0.001 37 EEGIGGTIPH 0.001 30 WEKSQFLEEG 0.001 29 GWEKSQFLEE 0.000 36 LEEGIGGTIP 0.000

TABLE XIX-V7A HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 2 SPKSLSETFL 80.000 6 LSETFLPNGI 0.120 9 TFLPNGINGI 0.040 1 GSPKSLSETF 0.020 4 KSLSETFLPN 0.020 10 FLPNGINGIK 0.010 5 SLSETFLPNG 0.010 8 ETFLPNGING 0.010 7 SETFLPNGIN 0.003 3 PKSLSETFLP 0.000

TABLE XIX-V7B HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 3 LNMAYQQSTL 12.000 8 QQSTLGYVAL 4.000 9 QSTLGYVALL 4.000 10 STLGYVALLI 0.400 7 YQQSTLGYVA 0.100 2 FLNMAYQQST 0.100 6 AYQQSTLGYV 0.060 5 MAYQQSTLGY 0.060 4 NMAYQQSTLG 0.010 1 LFLNMAYQQS 0.002

TABLE XIX-V7C HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 102 DPPESPDRAL 120.000 122 LPHTNGVGPL 80.000 129 GPLWEFLLRL 80.000 113 AANSWRNPVL 36.000 127 GVGPLWEFLL 20.000 15 ASPAAAWKCL 12.000 24 LGANILRGGL 6.000 152 FTSWSLGEFL 4.000 42 WQQDRKIPPL 4.000 160 FLGSGTWMKL 4.000 5 VILDLSVEVL 4.000 126 NGVGPLWEFL 4.000 141 SQAASGTLSL 4.000 119 NPVLPHTNGV 4.000 148 LSLAFTSWSL 4.000 19 AAWKCLGANI 3.600 28 ILRGGLSEIV 2.000 168 KLETIILSKL 1.200 20 AWKCLGANIL 1.200 165 TWMKLETIIL 1.200 66 ATAEAQESGI 1.200 4 IVILDLSVEV 1.000 135 LLRLLKSQAA 1.000 112 KAANSWRNPV 0.900 164 GTWMKLETII 0.400 139 LKSQAASGTL 0.400 181 QKSKHCMFSL 0.400 76 RNKSSSSSQI 0.400 29 LRGGLSEIVL 0.400 1 LPSIVILDLS 0.400 130 PLWEFLLRLL 0.400 27 NILRGGLSEI 0.400 31 GGLSEIVLPI 0.400 163 SGTWMKLETI 0.400 182 KSKHCMFSLI 0.400 144 ASGTLSLAFT 0.300 49 PPLSTPPPPA 0.300 81 SSSQIPVVGV 0.300 142 QAASGTLSLA 0.300 14 LASPAAAWKC 0.300 58 AMWTEEAGAT 0.300 178 TQEQKSKHCM 0.300 16 SPAAAWKCLG 0.200 85 IPVVGVVTED 0.200 82 SSQIPVVGVV 0.200 48 IPPLSTPPPP 0.200 55 PPPAMWTEEA 0.200 78 KSSSSSQIPV 0.200 79 SSSSSQIPVV 0.200 74 GIRNKSSSSS 0.200 53 TPPPPAMWTE 0.200 38 LPIEWQQDRK 0.200 18 AAAWKCLGAN 0.180 143 AASGTLSLAF 0.180 50 PLSTPPPPAM 0.150 10 SVEVLASPAA 0.150 52 STPPPPAMWT 0.150 44 QDRKIPPLST 0.150 12 EVLASPAAAW 0.150 106 SPDRALKAAN 0.120 158 GEFLGSGTWM 0.100 156 SLGEFLGSGT 0.100 162 GSGTWMKLET 0.100 88 VGVVTEDDEA 0.100 134 FLLRLLKSQA 0.100 138 LLKSQAASGT 0.100 177 LTQEQKSKHC 0.100 83 SQIPVVGVVT 0.100 105 ESPDRALKAA 0.100 116 SWRNPVLPHT 0.100 9 LSVEVLASPA 0.100 57 PAMWTEEAGA 0.090 185 HCMFSLISGS 0.060 110 ALKAANSWRN 0.060 25 GANILRGGLS 0.060 64 AGATAEAQES 0.060 36 IVLPIEWQQD 0.050 87 VVGVVTEDDE 0.050 90 VVTEDDEAQD 0.050 89 GVVTEDDEAQ 0.050 150 LAFTSWSLGE 0.030 125 TNGVGPLWEF 0.030 109 RALKAANSWR 0.030 96 EAQDSIDPPE 0.030 63 EAGATAEAQE 0.030 26 ANILRGGLSE 0.030 51 LSTPPPPAMW 0.030 69 EAQESGIRNK 0.030 17 PAAAWKCLGA 0.030 65 GATAEAQESG 0.030 114 ANSWRNPVLP 0.030 6 ILDLSVEVLA 0.030 70 AQESGIRNKS 0.027 147 TLSLAFTSWS 0.020 146 GTLSLAFTSW 0.020 140 KSQAASGTLS 0.020 180 EQKSKHCMFS 0.020 56 PPAMWTEEAG 0.020 145 SGTLSLAFTS 0.020 72 ESGIRNKSSS 0.020

TABLE XX-V1 HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 62 NPKFASEFF 60.000 323 LPMRRSERY 40.000 157 GPKDASRQV 24.000 259 LPIVAITLL 20.000 428 TPPNFVLAL 20.000 291 FPPWLETWL 20.000 438 LPSIVILDL 20.000 214 GPVVVAISL 20.000 231 FVRDVIHPY 12.000 37 GSGDFAKSL 10.000 405 ISTFHVLIY 10.000 204 LPLRLFTLW 10.000 239 YARNQQSDF 9.000 41 FAKSLTIRL 9.000 173 QARQQVIEL 9.000 99 TSLWDLRHL 7.500 196 SSAREIENL 7.500 9 SPKSLSETC 6.000 317 VAYSLCLPM 6.000 276 LAAAYQLYY 6.000 272 LAGLLAAAY 6.000 125 YPESNAEYL 6.000 46 TIRLIRCGY 6.000 267 LSLVYLAGL 5.000 395 QSTLGYVAL 5.000 366 MSLGLLSLL 5.000 220 ISLATFFFL 5.000 250 IPIEIVNKT 4.000 112 ILIDVSNNM 4.000 188 IPIDLGSLS 4.000 347 NSWNEEEVW 3.750 133 LASLFPDSL 3.000 300 QCRKQLGLL 3.000 218 VAISLATFF 3.000 177 QVIELARQL 2.000 303 KQLGLLSFF 2.000 371 LSLLAVTSI 2.000 128 SNAEYLASL 2.000 275 LLAAAYQLY 2.000 61 RNPKFASEF 2.000 100 SLWDLRHLL 2.000 237 HPYARNQQS 2.000 379 IPSVSNALN 2.000 117 SNNMRINQY 2.000 306 GLLSFFFAM 2.000 134 ASLFPDSLI 2.000 221 SLATFFFLY 2.000 193 GSLSSAREI 2.000 263 AITLLSLVY 2.000 90 FVAIHREHY 2.000 280 YQLYYGTKY 2.000 358 EMYISFGIM 2.000 27 DARKVTVGV 1.800 441 IVILDLLQL 1.500 161 ASRQVYICS 1.500 59 FIPIDLGSL 1.500 81 DALTKTNII 1.200 65 FASEFFPHV 1.200 365 IMSLGLLSL 1.000 184 QLNFIPIDL 1.000 385 ALNWREFSF 1.000 148 VVSAWALQL 1.000 274 GLLAAAYQL 1.000 144 KGFNVVSAW 1.000 146 FNVVSAWAL 1.000 383 SNALNWREF 1.000 304 QLGLLSFFF 1.000 363 FGIMSLGLL 1.000 217 VVAISLATF 1.000 57 VIGSRNPKF 1.000 313 AMVHVAYSL 1.000 411 LIYGWKRAF 1.000 378 SIPSVSNAL 1.000 264 ITLLSLVYL 1.000 75 DVTHHEDAL 1.000 436 LVLPSIVIL 1.000 82 ALTKTNIIF 1.000 403 LLISTFHVL 1.000 299 LQCRKQLGL 1.000 400 YVALLISTF 1.000 258 TLPIVAITL 1.000 268 SLVYLAGLL 1.000 5 SMMGSPKSL 1.000 223 ATEFFLYSF 1.000 33 VGVIGSGDF 1.000 396 STLGYVALL 1.000 261 IVAITLLSL 1.000 360 YISFGIMSL 1.000 219 AISLATFFF 1.000 203 NLPLRLFTL 1.000 129 NAEYLASLF 0.900 85 KTNIIFVAI 0.800 127 ESNAEYLAS 0.750 386 LNWREFSFI 0.600 434 LALVLPSIV 0.600 416 KRAFEEEYY 0.600 328 SERYLFLNM 0.600 28 KYRRFPPWL 0.600 24 GIKDARKVT 0.600

TABLE XX-V2 HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 37 CPADFFLYF 40.000 33 CPPPCPADF 20.000 3 SPGLQALSL 20.000 23 LSLPSSWDY 10.000 9 LSLSLSSGF 5.000 35 PPCPADFFL 2.000 34 PPPCPADFF 2.000 25 LPSSWDYRC 2.000 15 SGFTPFSCL 1.000 1 SGSPGLQAL 1.000 12 SLSSGFTPF 1.000 5 GLQALSLSL 1.000 17 FTPFSCLSL 1.000 20 FSCLSLPSS 0.500 2 GSPGLQALS 0.500 13 LSSGFTPFS 0.500 14 SSGFTPFSC 0.500 21 SCLSLPSSW 0.500 7 QALSLSLSS 0.300 36 PCPADFFLY 0.300 18 TPFSCLSLP 0.200 6 LQALSLSLS 0.100 10 SLSLSSGFT 0.100 27 SSWDYRCPP 0.100 11 LSLSSGFTP 0.050 32 RCPPPCPAD 0.020 8 ALSLSLSSG 0.010 22 CLSLPSSWD 0.010 29 WDYRCPPPC 0.010 24 SLPSSWDYR 0.010 31 YRCPPPCPA 0.010 4 PGLQALSLS 0.010 16 GFTPFSCLS 0.010 26 PSSWDYRCP 0.008 30 DYRCPPPCP 0.003 19 PFSCLSLPS 0.001 28 SWDYRCPPP 0.000

TABLE XX-V5A HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 2 LPLRLFTFW 10.000 1 NLPLRLFTF 1.000 7 FTFWRGPVV 0.200 9 FWRGPVVVA 0.030 6 LFTFWRGPV 0.020 5 RLFTFWRGP 0.020 8 TFWRGPVVV 0.020 3 PLRLFTFWR 0.003 4 LRLFTFWRG 0.001

TABLE XX-V5B HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 16 TQTELELEF 2.000 24 FVFLLTLLL 1.000 4 FSFIQIFCS 0.500 19 ELELEFVFL 0.450 12 SFADTQTEL 0.200 18 TELELEFVF 0.200 20 LELEFVFLL 0.200 2 REESFIQIF 0.200 22 LEFVFLLTL 0.100 10 FCSFADTQT 0.100 8 QIFCSFADT 0.100 23 EFVFLLTLL 0.100 6 FIQIFCSFA 0.100 14 ADTQTELEL 0.100 5 SFIQIFCSF 0.100 17 QTELELEFV 0.090 11 CSFADTQTE 0.075 21 ELEFVFLLT 0.030 15 DTQTELELE 0.015 1 WREFSFIQI 0.012 7 IQIFCSFAD 0.010 3 EFSFIQIFC 0.010 13 FADTQTELE 0.009 9 IFCSFADTQ 0.001

TABLE XX-V6 HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 46 VSPERVTVM 20.000 27 KGWEKSQFL 4.000 43 IPHVSPERV 4.000 31 KSQFLEEGI 4.000 21 KLKRIKKGW 3.000 14 FLPCISRKL 1.000 6 VILGKIILF 1.000 5 IVILGLIIL 1.000 7 ILGKIILFL 1.000 10 KIILFLPCI 0.800 24 RIKKGWEKS 0.600 17 CISRKLKRI 0.400 4 SIVILGKII 0.400 45 HVSPERVTV 0.300 26 KKGWEKSQF 0.300 2 LPSIVILGK 0.200 15 LPCISRKLK 0.200 38 GIGGTIPHV 0.200 3 PSIVILGKI 0.200 18 ISRKLKRIK 0.150 39 IGGTIPHVS 0.100 11 IILFLPCIS 0.100 34 FLEEGIGGT 0.060 8 LGKIILFLP 0.030 32 SQFLEEGIG 0.015 35 LEEGIGGTI 0.012 37 EGIGGTIPH 0.010 41 GTIPHVSPE 0.010 40 GGTIPHVSP 0.010 1 VLPSIVILG 0.010 9 GKIILFLPC 0.010 12 ILFLPCISR 0.010 42 TIPHVSPER 0.010 33 QFLEEGIGG 0.003 29 WEKSQFLEE 0.003 25 IKKGWEKSQ 0.003 22 LKRIKKGWE 0.003 19 SRKLKRIKK 0.003 20 RKLKRIKKG 0.002 23 KRIKKGWEK 0.002 44 PHVSPERVT 0.001 13 LFLPCISRK 0.001 30 EKSQFLEEG 0.001 16 PCISRKLKR 0.001 36 EEGIGGTIP 0.001 28 GWEKSQFLE 0.000

TABLE XX-V7A HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 SPKSLSETF 60.000 9 FLPNGINGI 0.400 4 SLSETFLPN 0.200 3 KSLSETFLP 0.150 7 ETFLPNGIN 0.100 6 SETFLPNGI 0.040 5 LSETFLPNG 0.015 2 PKSLSETFL 0.010 8 TFLPNGING 0.001

TABLE XX-V7B HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 8 QSTLGYVAL 5.000 9 STLGYVALL 1.000 3 NMAYQQSTL 1.000 6 YQQSTLGYV 0.200 5 AYQQSTLGY 0.200 7 QQSTLGYVA 0.100 1 FLNMAYQQS 0.100 2 LNMAYQQST 0.100 4 MAYQQSTLG 0.030

TABLE XX-V7C HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 181 KSKHCMFSL 30.000 15 SPAAAWKCL 20.000 139 KSQAASGTL 10.000 50 LSTPPPPAM 10.000 152 TSWSLGEFL 5.000 143 ASGTLSLAF 5.000 165 WMKLETIIL 4.500 101 DPPESPDRA 4.000 179 EQKSKHCMF 3.000 24 GANILRGGL 3.000 141 QAASGTLSL 3.000 108 RALKAANSW 3.000 29 RGGLSEIVL 2.000 52 TPPPPAMWT 2.000 27 ILRGGLSEI 1.200 78 SSSSSQIPV 1.000 126 GVGPLWEFL 1.000 113 ANSWRNPVL 1.000 104 ESPDRALKA 1.000 160 LGSGTWMKL 1.000 127 VGPLWEFLL 1.000 79 SSSSQIPVV 1.000 148 SLAFTSWSL 1.000 151 FTSWSLGEF 1.000 125 NGVGPLWEF 1.000 81 SSQIPVVGV 1.000 31 GLSEIVLPI 0.800 154 WSLGEFLGS 0.750 102 PPESPDRAL 0.600 112 AANSWRNPV 0.600 105 SPDRALKAA 0.600 68 EAQESGIRN 0.600 51 STPPPPAMW 0.500 147 LSLAFTSWS 0.500 146 TLSLAFTSW 0.500 12 VLASPAAAW 0.500 71 ESGIRNKSS 0.500 123 HTNGVGPLW 0.500 14 ASPAAAWKC 0.500 64 GATAEAQES 0.450 163 GTWMKLETI 0.400 37 LPIEWQQDR 0.400 4 VILDLSVEV 0.400 66 TAEAQESGI 0.360 134 LLRLLKSQA 0.300 42 QQDRKIPPL 0.300 73 GIRNKSSSS 0.300 17 PAAWKCLGA 0.300 142 AASGTLSLA 0.300 128 GPLWEFLLR 0.300 18 AAWKCLGAN 0.300 5 ILDLSVEVL 0.300 136 RLLKSQAAS 0.200 82 SQIPVVGVV 0.200 47 IPPLSTPPP 0.200 55 PPAMWTEEA 0.200 121 LPHTNGVGP 0.200 129 PLWEFLLRL 0.200 178 QEQKSKHCM 0.200 117 RNPVLPHTN 0.200 2 SIVILDLSV 0.200 158 EFLGSGTWM 0.200 84 IPVVGVVTE 0.200 118 NPVLPHTNG 0.200 57 AMWTEEAGA 0.150 173 LSKLTQEQK 0.150 7 DLSVEVLAS 0.150 88 GVVTEDDEA 0.150 19 AWKCLGANI 0.120 98 DSIDPPESP 0.100 145 GTLSLAFTS 0.100 83 QIPVVGVVT 0.100 8 LSVEVLASP 0.100 168 LETIILSKL 0.100 169 ETIILSKLT 0.100 162 SGTWMKLET 0.100 11 EVLASPAAA 0.100 25 ANILRGGLS 0.100 72 SGIRNKSSS 0.100 144 SGTLSLAFT 0.100 140 SQAASGTLS 0.100 77 KSSSSSQIP 0.100 185 CMFSLISGS 0.100 20 WKCLGANIL 0.100 95 EAQDSIDPP 0.060 111 KAANSWRNP 0.060 75 RNKSSSSSQ 0.060 59 WTEEAGATA 0.060 1 PSIVILDLS 0.050 80 SSSQIPVVG 0.050 157 GEFLGSGTW 0.050 33 SEIVLPIEW 0.050 161 GSGTWMKLE 0.050 114 NSWRNPVLP 0.050 76 NKSSSSSQI 0.040 164 TWMKLETII 0.040 182 SKHCMFSLI 0.040 39 IEWQQDRKI 0.040 58 MWTEEAGAT 0.030 89 WTEDDEAQ 0.030

TABLE XXI-V1 HLA-B3501-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 157 GPKDASRQVY 240.000 9 SPKSLSETCL 60.000 250 IPIEIVNKTL 40.000 197 SAREIENLPL 27.000 323 LPMRRSERYL 20.000 438 LPSIVILDLL 20.000 239 YARNQQSDFY 18.000 417 RAFEEEYYRF 18.000 379 IPSVSNALNW 10.000 116 VSNNMRINQY 10.000 391 FSFIQSTLGY 10.000 220 ISLATFFFLY 10.000 195 LSSAREIENL 7.500 137 FPDSLIVKGF 6.000 327 RSERYLFLNM 6.000 262 VAITLLSLVY 6.000 361 ISFGIMSLGL 5.000 395 QSTLGYVALL 5.000 267 LSLVYLAGLL 5.000 99 TSLWDLRHLL 5.000 127 ESNAEYLASL 5.000 4 ISMMGSPKSL 5.000 382 VSNALNWREF 5.000 377 TSIPSVSNAL 5.000 428 TPPNFVLALV 4.000 188 IPIDLGSLSS 4.000 111 KILIDVSNNM 4.000 181 LARQLNFIPI 3.600 27 DARKVTVGVI 3.600 41 FAKSLTIRLI 3.600 384 NALNWREFSF 3.000 312 FAMVHVAYSL 3.000 222 LATFFFLYSF 3.000 81 DALTKTNIIF 3.000 218 VAISLATFFF 3.000 322 CLPMRRSERY 2.000 429 PPNFVLALVL 2.000 316 HVAYSLCLPM 2.000 61 RNPKFASEFF 2.000 257 KTLPIVAITL 2.000 259 LPIVAITLLS 2.000 45 LTIRLIRCGY 2.000 275 LLAAAYQLYY 2.000 274 GLLAAAYQLY 2.000 303 KQLGLLSFFF 2.000 128 SNAEYLASLF 2.000 123 NQYPESNAEY 2.000 305 LGLLSFFFAM 2.000 404 LISTFHVLIY 2.000 213 RGPVVVAISL 2.000 271 YLAGLLAAAY 2.000 183 RQLNFIPIDL 2.000 214 GPVVVAISLA 2.000 134 ASLFPDSLIV 1.500 440 SIVILDLLQL 1.500 98 YTSLWDLRHL 1.500 161 ASRQVYICSN 1.500 285 GTKYRRFPPW 1.500 103 DLRHLLVGKI 1.200 336 MAYQQVHANI 1.200 255 VNKTLPIVAI 1.200 65 FASEFFPHVV 1.200 49 LIRCGYHVVI 1.200 434 LALVLPSIVI 1.200 133 LASLFPDSLI 1.200 24 GIKDARKVTV 1.200 241 RNQQSDFYKI 1.200 32 TVGVIGSGDF 1.000 435 ALVLPSIVIL 1.000 273 AGLLAAAYQL 1.000 36 IGSGDFAKSL 1.000 308 LSFFFAMVHV 1.000 56 VVIGSRNPKF 1.000 176 QQVIELARQL 1.000 296 ETWLQCRKQL 1.000 43 KSLTIRLIRC 1.000 202 ENLPLRLFTL 1.000 147 NVVSAWALQL 1.000 217 VVAISLATFF 1.000 216 VVVAISLATF 1.000 132 YLASLFPDSL 1.000 364 GIMSLGLLSL 1.000 365 IMSLGLLSLL 1.000 92 AIHREHYTSL 1.000 314 MVHVAYSLCL 1.000 410 VLIYGWKRAF 1.000 299 LQCRKQLGLL 1.000 394 IQSTLGYVAL 1.000 11 KSLSETCLPN 1.000 263 AITLLSLVYL 1.000 172 IQARQQVIEL 1.000 219 AISLATFFFL 1.000 298 WLQCRKQLGL 1.000 37 GSGDFAKSLT 1.000 402 ALLISTFHVL 1.000 258 TLPIVAITLL 1.000 427 YTPPNFVLAL 1.000 139 DSLIVKGFNV 1.000 437 VLPSIVILDL 1.000 266 LLSLVYLAGL 1.000

TABLE XXI-V2 HLA-B3501-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 33 CPPPCPADFF 20.000 35 PPCPADFFLY 6.000 14 SSGFTPFSCL 5.000 11 LSLSSGFTPF 5.000 2 GSPGLQALSL 5.000 20 FSCLSLPSSW 2.500 34 PPPCPADFFL 2.000 3 SPGLQALSLS 2.000 22 CLSLPSSWDY 2.000 32 RCPPPCPADF 2.000 18 TPFSCLSLPS 2.000 8 ALSLSLSSGF 1.000 9 LSLSLSSGFT 0.500 13 LSSGFTPFSC 0.500 25 LPSSWDYRCP 0.300 4 PGLQALSLSL 0.100 15 SGFTPFSCLS 0.100 27 SSWDYRCPPP 0.100 16 GFTPFSCLSL 0.100 6 LQALSLSLSS 0.100 1 SGSPGLQALS 0.100 24 SLPSSWDYRC 0.100 36 PCPADFFLYF 0.100 5 GLQALSLSLS 0.100 12 SLSSGFTPFS 0.100 23 LSLPSSWDYR 0.050 7 QALSLSLSSG 0.030 30 DYRCPPPCPA 0.030 17 FTPFSCLSLP 0.010 10 SLSLSSGFTP 0.010 21 SCLSLPSSWD 0.010 26 PSSWDYRCPP 0.005 28 SWDYRCPPPC 0.003 29 WDYRCPPPCP 0.001 19 PFSCLSLPSS 0.001 31 YRCPPPCPAD 0.001

TABLE XXI-V5A HLA-B3501-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 1 ENLPLRLFTF 1.000 2 NLPLRLFTFW 0.500 6 RLFTFWRGPV 0.400 8 FTFWRGPVVV 0.200 3 LPLRLFTFWR 0.200 10 FWRGPVVVAI 0.120 7 LFTFWRGPVV 0.020 9 TFWRGPVVVA 0.010 4 PLRLFTFWRG 0.003 5 LRLFTFWRGP 0.001

TABLE XXI-V5B HLA-3501-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 12 CSFADTQTEL 5.000 5 FSFIQIFCSF 5.000 16 DTQTELELEF 1.000 14 FADTQTELEL 0.900 17 TQTELELEFV 0.600 22 ELEFVFLLTL 0.300 18 QTELELEFVF 0.300 20 ELELEFVFLL 0.300 19 TELELEFVFL 0.300 1 NWREFSFIQI 0.240 8 IQIFCSFADT 0.100 23 LEFVFLLTLL 0.100 24 EFVFLLTLLL 0.100 2 WREFSFIQIF 0.030 3 REFSFIQIFC 0.020 21 LELEFVFLLT 0.020 11 FCSFADTQTE 0.015 10 IFCSFADTQT 0.010 7 FIQIFCSFAD 0.010 4 EFSFIQIFCS 0.010 9 QIFCSFADTQ 0.010 6 SFIQIFCSFA 0.010 13 SFADTQTELE 0.002 15 ADTQTELELE 0.002

TABLE XXI-V6 HLA-B3501-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 3 LPSIVILGKI 8.000 44 IPHVSPERVT 2.000 46 HVSPERVTVM 2.000 6 IVILGKIILF 1.000 7 VILGKIILFL 1.000 5 SIVILGKILL 1.000 26 IKKGWEKSQF 0.450 9 LGKIILFLPC 0.300 35 FLEEGIGGTI 0.240 43 TIPHVSPERV 0.200 11 KIILFLPCIS 0.200 27 KKGWEKSQFL 0.200 38 EGIGGTIPHV 0.200 16 LPCISRKLKR 0.200 4 PSIVILGKII 0.200 32 KSQFLEEGIG 0.150 19 ISRKLKRIKK 0.150 39 GIGGTIPHVS 0.100 14 LFLPCISRKL 0.100 21 RKLKRIKKGW 0.100 25 RIKKGWEKSQ 0.060 22 KLKRIKKGWE 0.060 10 GKIILFLPCI 0.040 28 KGWEKSQFLE 0.040 17 PCISRKLKRI 0.040 31 EKSQFLEEGI 0.040 24 KRIKKGWEKS 0.020 34 QFLEEGIGGT 0.020 33 SQFLEEGIGG 0.015 13 ILFLPCISRK 0.010 18 CISRKLKRIK 0.010 8 ILGKIILFLP 0.010 2 VLPSIVILGK 0.010 40 IGGTIPHVSP 0.010 15 FLPCISRKLK 0.010 41 GGTIPHVSPE 0.010 1 LVLPSIVILG 0.010 42 GTIPHVSPER 0.010 12 IILFLPCISR 0.010 45 PHVSPERVTV 0.003 20 SRKLKRIKKG 0.003 30 WEKSQFLEEG 0.003 23 LKRIKKGWEK 0.003 37 EEGIGGTIPH 0.001 36 LEEGIGGTIP 0.000 29 GWEKSQFLEE 0.000

TABLE XXI-V7A HLA-3501-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 2 SPKSLSETFL 60.000 1 GSPKSLSETF 5.000 4 KSLSETFLPN 1.000 6 LSETFLPNGI 0.600 9 TFLPNGINGI 0.040 5 SLSETFLPNG 0.020 10 FLPNGINGIK 0.010 7 SETFLPNGIN 0.010 8 ETFLPNGING 0.010 3 PKSLSETFLP 0.000

TABLE XXI-V7B HLA-B3501-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 MAYQQSTLGY 6.000 9 QSTLGYVALL 5.000 3 LNMAYQQSTL 1.000 8 QQSTLGYVAL 1.000 10 STLGYVALLI 0.400 7 YQQSTLGYVA 0.100 2 FLNMAYQQST 0.100 6 AYQQSTLGYV 0.020 4 NMAYQQSTLG 0.010 1 LFLNMAYQQS 0.010

TABLE XXI-V7C HLA-B3501-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 100 SIDPPESPDR 100.000 67 TAEAQESGIR 9.000 33 LSEIVLPIEW 6.750 131 LWEFLLRLLK 4.500 91 VTEDDEAQDS 2.250 10 SVEVLASPAA 1.800 52 STPPPPAMWT 1.250 6 ILDLSVEVLA 1.000 168 KLETIILSKL 0.900 103 PPESPDRALK 0.900 127 GVGPLWEFLL 0.500 143 AASGTLSLAF 0.500 13 VLASPAAAWK 0.400 51 LSTPPPPAMW 0.300 60 WTEEAGATAE 0.225 157 LGEFLGSGTW 0.225 69 EAQESGIRNK 0.200 97 AQDSIDPPES 0.150 70 AQESGIRNKS 0.135 178 TQEQKSKHCM 0.135 170 ETIILSKLTQ 0.125 128 VGPLWEFLLR 0.125 37 VLPIEWQQDR 0.100 14 LASPAAAWKC 0.100 61 TEEAGATAEA 0.090 39 PIEWQQDRKI 0.090 162 GSGTWMKLET 0.075 78 KSSSSSQIPV 0.075 160 FLGSGTWMKL 0.050 22 KCLGANILRG 0.050 167 MKLETIILSK 0.050 38 LPIEWQQDRK 0.050 80 SSSSQIPVVG 0.030 79 SSSSSQIPVV 0.030 83 SQIPVVGVVT 0.030 144 ASGTLSLAFT 0.030 81 SSSQIPVVGV 0.030 146 GTLSLAFTSW 0.025 66 ATAEAQESGI 0.025 152 FTSWSLGEFL 0.025 125 TNGVGPLWEF 0.025 92 TEDDEAQDSI 0.025 177 LTQEQKSKHC 0.025 21 WKCLGANILR 0.025 106 SPDRALKAAN 0.025 94 DDEAQDSIDP 0.022 12 EVLASPAAAW 0.020 4 IVILDLSVEV 0.020 173 ILSKLTQEQK 0.020 47 KIPPLSTPPP 0.020 113 AANSWRNPVL 0.020 72 ESGIRNKSSS 0.015 43 QQDRKIPPLS 0.015 15 ASPAAAWKCL 0.015 140 KSQAASGTLS 0.015 9 LSVEVLASPA 0.015 82 SSQIPVVGVV 0.015 155 WSLGEFLGSG 0.015 105 ESPDRALKAA 0.015 148 LSLAFTSWSL 0.015 124 HTNGVGPLWE 0.013 129 GPLWEFLLRL 0.013 31 GGLSEIVLPI 0.013 145 SGTLSLAFTS 0.013 185 HCMFSLISGS 0.010 149 SLAFTSWSLG 0.010 65 GATAEAQESG 0.010 112 KAANSWRNPV 0.010 142 QAASGTLSLA 0.010 25 GANILRGGLS 0.010 159 EFLGSGTWMK 0.010 23 CLGANILRGG 0.010 109 RALKAANSWR 0.010 176 KLTQEQKSKH 0.010 35 EIVLPIEWQQ 0.010 175 SKLTQEQKSK 0.010 18 AAAWKCLGAN 0.010 36 IVLPIEWQQD 0.010 5 VILDLSVEVL 0.010 172 IILSKLTQEQ 0.010 156 SLGEFLGSGT 0.010 120 PVLPHTNGVG 0.010 147 TLSLAFTSWS 0.010 89 GVVTEDDEAQ 0.010 153 TSWSLGEFLG 0.008 2 PSIVILDLSV 0.008 141 SQAASGTLSL 0.007 150 LAFTSWSLGE 0.005 17 PAAAWKCLGA 0.005 101 IDPPESPDRA 0.005 151 AFTSWSLGEF 0.005 117 WRNPVLPHTN 0.005 42 WQQDRKIPPL 0.003 104 PESPDRALKA 0.003 24 LGANILRGGL 0.003 119 NPVLPHTNGV 0.003 118 RNPVLPHTNG 0.003 102 DPPESPDRAL 0.003 53 TPPPPAMWTE 0.003 1 LPSIVILDLS 0.003

TABLE VIII-V8 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 4 FLEEGMGGT 0.900 5 LEEGMGGTI 0.045 1 KSQFLEEGM 0.015 7 EGMGGTIPH 0.013 8 GMGGTIPHV 0.010 9 MGGTIPHVS 0.003 3 QFLEEGMGG 0.003 2 SQFLEEGMG 0.002 6 EEGMGGTIP 0.000

TABLE VIII-V13 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 5 LSETFLPNG 2.700 4 SLSETFLPN 0.050 7 ETFLPNGIN 0.025 8 TFLPNGING 0.025 9 FLPNGINGI 0.010 3 KSLSETFLP 0.007 1 SPKSLSETF 0.003 6 SETFLPNGI 0.001 2 PKSLSETFL 0.000

TABLE VIII-V14 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 NLPLRLFTF 0.500 7 FTFWRGPVV 0.050 3 PLRLFTFWR 0.005 5 RLFTFWRGP 0.001 6 LFTFWRGPV 0.001 4 LRLFTFWRG 0.001 2 LPLRLFTFW 0.000 9 FWRGPVVVA 0.000 8 TFWRGPVVV 0.000

TABLE VIII-V21 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 2 KLTQEQKTK 0.200 4 TQEQKTKHC 0.135 3 LTQEQKTKH 0.025 8 KTKHCMFSL 0.013 6 EQKTKHCMF 0.002 9 TKHCMFSLI 0.001 1 SKLTQEQKT 0.001 7 QKTKHCMFS 0.000 5 QEQKTKHCM 0.000

TABLE VIII-V25 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 2 LFLPCISQK 0.100 1 ILFLPCISQ 0.050 5 PCISQKLKR 0.050 4 LPCISQKLK 0.050 7 ISQKLKRIK 0.030 8 SQKLKRIKK 0.015 3 FLPCISQKL 0.010 6 CISQKLKRI 0.010 9 QKLKRIKKG 0.000

TABLE IX-V8 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 FLEEGMGGTI 0.900 2 KSQFLEEGMG 0.015 3 SQFLEEGMGG 0.007 8 EGMGGTIPHV 0.005 9 GMGGTIPHVS 0.005 6 LEEGMGGTIP 0.005 7 EEGMGGTIPH 0.003 4 QFLEEGMGGT 0.001 10 MGGTIPHVSP 0.001 1 EKSQFLEEGM 0.001

TABLE IX-V13 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 6 LSETFLPNGI 1.350 10 FLPNGINGIK 0.200 8 ETFLPNGING 0.125 4 KSLSETFLPN 0.075 5 SLSETFLPNG 0.020 1 GSPKSLSETF 0.015 9 TFLPNGINGI 0.005 7 SETFLPNGIN 0.001 2 SPKSLSETFL 0.000 3 PKSLSETFLP 0.000

TABLE IX-V14 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 1 ENLPLRLFTF 1.250 8 FTFWRGPVVV 0.050 3 LPLRLFTFWR 0.013 2 NLPLRLFTFW 0.010 6 RLFTFWRGPV 0.010 7 LFTFWRGPVV 0.001 4 PLRLFTFWRG 0.000 10 FWRGPVVVAI 0.000 5 LRLFTFWRGP 0.000 9 TFWRGPVVVA 0.000

TABLE IX-V21 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 TQEQKTKHCM 0.135 4 LTQEQKTKHC 0.025 3 KLTQEQKTKH 0.010 2 SKLTQEQKTK 0.010 9 KTKHCMFSLI 0.003 10 TKHCMFSLIS 0.003 1 LSKLTQEQKT 0.002 7 EQKTKHCMFS 0.001 6 QEQKTKHCMF 0.001 8 QKTKHCMFSL 0.000

TABLE IX-V25 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine. Start Subsequence Score 7 CISQKLKRIK 0.200 4 FLPCISQKLK 0.200 2 ILFLPCISQK 0.200 8 ISQKLKRIKK 0.150 5 LPCISQKLKR 0.125 1 IILFLPCISQ 0.050 3 LFLPCISQKL 0.005 6 PCISQKLKRI 0.001 9 SQKLKRIKKG 0.000 10 QKLKRIKKGW 0.000

TABLE X-V8 A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 8 GMGGTIPHV 115.534 4 FLEEGMGGT 2.689 1 KSQFLEEGM 0.056 2 SQFLEEGMG 0.004 5 LEEGMGGTI 0.003 3 QFLEEGMGG 0.001 9 MGGTIPHVS 0.000 7 EGMGGTIPH 0.000 6 EEGMGGTIP 0.000

TABLE X-V13 A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 FLPNGINGI 110.379 4 SLSETFLPN 0.581 6 SETFLPNGI 0.203 3 KSLSETFLP 0.007 2 PKSLSETFL 0.004 5 LSETFLPNG 0.000 8 TFLPNGING 0.000 7 ETFLPNGIN 0.000 1 SPKSLSETF 0.000

TABLE X-V14 A0201.9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 7 FTFWRGPVV 6.741 1 NLPLRLFTF 0.994 8 TFWRGPVVV 0.164 5 RLFTFWRGP 0.071 2 LPLRLFTFW 0.032 6 LFTFWRGPV 0.011 3 PLRLFTFWR 0.003 4 LRLFTFWRG 0.001 9 FWRGPVVVA 0.000

TABLE X-V21 A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 8 KTKHCMFSL 0.485 5 QEQKTKHCM 0.097 1 KLTQEQKTK 0.052 1 SKLTQEQKT 0.038 4 TQEQKTKHC 0.032 9 TKHCMFSLI 0.028 3 LTQEQKTKH 0.007 7 QKTKHCMFS 0.001 6 EQKTKHCMF 0.000

TABLE X-V25 A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 3 FLPCISQKL 98.267 6 CISQKLKRI 3.299 1 ILFLPCISQ 0.094 9 QKLKRIKKG 0.001 4 KPCISQKLK 0.000 2 LFLPCISQK 0.000 8 SQKLKRIKK 0.000 7 ISQKLKRIK 0.000 5 PCISQKLKR 0.000

TABLE X-V8 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 FLEEGMGGTI 1.637 8 EGMGGTIPHV 0.290 3 SQFLEEGMGG 0.028 4 QFLEEGMGGT 0.023 9 GMGGTIPHVS 0.022 1 EKSQFLEEGM 0.000 2 KSQFLEEGMG 0.000 10 MGGTIPHVSP 0.000 7 EEGMGGTIPH 0.000 6 LEEGMGGTIP 0.000

TABLE X-V13 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 SLSETFLPNG 2.670 9 TFLPNGINGI 0.062 2 SPKSLSETFL 0.027 4 KSLSETFLPN 0.012 6 LSETFLPNGI 0.007 10 FLPNGINGIK 0.004 8 ETFLPNGING 0.000 1 GSPKSLSETF 0.000 7 SETFLPNGIN 0.000 3 PKSLSETFLP 0.000

TABLE X-V14 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine. Start Subsequence Score 6 RLFTFWRGPV 33.455 8 FTFWRGPVVV 6.741 2 NLPLRLFTFW 0.779 3 LPLRLFTFWR 0.074 7 LFTFWRGPVV 0.034 9 TFWRGPVVVA 0.027 1 ENLPLRLFTF 0.002 4 PLRLFTFWRG 0.002 10 FWRGPVVVAI 0.001 5 LRLFTFWRGP 0.000

TABLE X-V21 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 TQEQKTKHCM 0.135 4 LTQEQKTKHC 0.025 3 KLTQEQKTKH 0.010 2 SKLTQEQKTK 0.010 9 KTKHCMFSLI 0.003 10 TKHCMFSLIS 0.003 1 LSKLTQEQKT 0.002 7 EQKTKHCMFS 0.001 6 QEQKTKHCMF 0.001 8 QKTKHCMFSL 0.000

TABLE X-V25 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine. Start Subsequence Score 2 ILFLPCISQK 0.216 3 LFLPCISQKL 0.093 4 FLPCISQKLK 0.069 1 IILFLPCISQ 0.013 6 PCISQKLKRI 0.003 9 SQKLKRIKKG 0.001 10 QKLKRIKKGW 0.000 7 CISQKLKRIK 0.000 8 ISQKLKRIKK 0.000 5 LPCISQKLKR 0.000

TABLE XII-V8 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 8 GMGGTIPHV 1.350 4 FLEEGMGGT 0.068 1 KSQFLEEGM 0.003 2 SQFLEEGMG 0.001 5 LEEGMGGTI 0.001 7 EGMGGTIPH 0.000 3 QFLEEGMGG 0.000 9 MGGTIPHVS 0.000 6 EEGMGGTIP 0.000

TABLE XII-V13 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 FLPNGINGI 0.900 4 SLSETFLPN 0.180 1 SPKSLSETF 0.020 6 SETFLPNGI 0.002 3 KSLSETFLP 0.001 7 ETFLPNGIN 0.001 5 LSETFLPNG 0.000 8 TFLPNGING 0.000 2 PKSLSETFL 0.000

TABLE XII-V14 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 NLPLRLFTF 9.000 3 PLRLFTFWR 3.600 7 FTFWRGPVV 0.050 5 RLFTFWRGP 0.030 2 LPLRLFTFW 0.009 9 FWRGPVVVA 0.001 8 TFWRGPVVV 0.001 4 LRLFTFWRG 0.000 6 LFTFWRGPV 0.000

TABLE XII-V21 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 2 KLTQEQKTK 30.000 8 KTKHCMFSL 0.405 6 EQKTKHCMF 0.018 3 LTQEQKTKH 0.015 4 TQEQKTKHC 0.003 9 TKHCMFSLI 0.002 5 QEQKTKHCM 0.001 1 SKLTQEQKT 0.000 7 QKTKHCMFS 0.000

TABLE XII-V25 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 8 SQKLKRIKK 1.200 3 FLPCISQKL 0.900 1 ILFLPCISQ 0.300 4 LPCISQKLK 0.100 2 LFLPCISQK 0.068 6 CISQKLKRI 0.045 5 PCISQKLKR 0.012 7 ISQKLKRIK 0.010 9 QKLKRIKKG 0.000

TABLE XIII-V8 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 9 GMGGTIPHVS 0.270 5 FLEEGMGGTI 0.270 3 SQFLEEGMGG 0.006 7 EEGMGGTIPH 0.000 8 EGMGGTIPHV 0.000 4 QFLEEGMGGT 0.000 6 LEEGMGGTIP 0.000 2 KSQFLEEGMG 0.000 1 EKSQFLEEGM 0.000 10 MGGTIPHVSP 0.000

TABLE XIII-V13 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 10 FLPNGINGIK 9.000 5 SLSETFLPNG 0.135 1 GSPKSLSETF 0.030 2 SPKSLSETFL 0.006 6 LSETFLPNGI 0.003 8 ETFLPNGING 0.003 4 KSLSETFLPN 0.003 9 TFLPNGINGI 0.002 7 SETFLPNGIN 0.000 3 PKSLSETFLP 0.000

TABLE XIII-V14 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 6 RLFTFWRGPV 0.900 2 NLPLRLFTFW 0.600 3 LPLRLFTFWR 0.540 8 FTFWRGPVVV 0.050 4 PLRLFTFWRG 0.018 1 ENLPLRLFTF 0.012 9 TFWRGPVVVA 0.005 10 FWRGPVVVAI 0.005 7 LFTFWRGPVV 0.000 5 LRLFTFWRGP 0.000

TABLE XIII-V21 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 3 KLTQEQKTKH 0.600 9 KTKHCMFSLI 0.270 2 SKLTQEQKTK 0.015 4 LTQEQKTKHC 0.007 6 QEQKTKHCMF 0.006 5 TQEQKTKHCM 0.006 8 QKTKHCMFSL 0.003 7 EQKTKHCMFS 0.001 1 LSKLTQEQKT 0.001 10 TKHCMFSLIS 0.000

TABLE XIII-V25 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 2 ILFLPCISQK 150.000 4 FLPCISQKLK 10.000 8 ISQKLKRIKK 0.200 7 CISQKLKRIK 0.080 5 LPCISQKLKR 0.080 1 IILFLPCISQ 0.009 3 LFLPCISQKL 0.002 6 PCISQKLKRI 0.001 9 SQKLKRIKKG 0.000 10 QKLKRIKKGW 0.000

TABLE XIV-V8 HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 8 GMGGTIPHV 1.350 4 FLEEGMGGT 0.068 1 KSQFLEEGM 0.003 2 SQFLEEGMG 0.001 5 LEEGMGGTI 0.001 7 EGMGGTIPH 0.000 3 QFLEEGMGG 0.000 9 MGGTIPHVS 0.000 6 EEGMGGTIP 0.000

TABLE XIV-V13 HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 FLPNGINGI 0.004 1 SPKSLSETF 0.002 4 SLSETFLPN 0.001 7 ETFLPNGIN 0.001 8 TFLPNGING 0.001 6 SETFLPNGI 0.001 3 KSLSETFLP 0.000 2 PKSLSETFL 0.000 5 LSETFLPNG 0.000

TABLE XIV-V14 HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 3 PLRLFTFWR 0.024 7 FTFWRGPVV 0.020 1 NLPLRLFTF 0.012 8 TFWRGPVVV 0.004 2 LPLRLFTFW 0.003 6 LFTFWRGPV 0.002 5 RLFTFWRGP 0.000 9 FWRGPVVVA 0.000 4 LRLFTFWRG 0.000

TABLE XIV-V21 HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 2 KLTQEQKTK 0.600 8 KTKHCMFSL 0.090 3 LTQEQKTKH 0.010 6 EQKTKHCMF 0.002 5 QEQKTKHCM 0.001 4 TQEQKTKHC 0.000 9 TKHCMFSLI 0.000 7 QKTKHCMFS 0.000 1 SKLTQEQKT 0.000

TABLE XIV-V25 HLA-A1101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 8 SQKLKRIKK 1.200 2 LFLPCISQK 0.300 4 LPCISQKLK 0.100 5 PCISQKLKR 0.012 3 FLPCISQKL 0.004 7 ISQKLKRIK 0.002 6 CISQKLKRI 0.002 1 ILFLPCISQ 0.002 9 QKLKRIKKG 0.000

TABLE XV-V8 HLA-A11-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 FLEEGMGGTI 0.004 3 SQFLEEGMGG 0.002 9 GMGGTIPHVS 0.001 7 EEGMGGTIPH 0.000 4 QFLEEGMGGT 0.000 8 EGMGGTIPHV 0.000 2 KSQFLEEGMG 0.000 6 LEEGMGGTIP 0.000 1 EKSQFLEEGM 0.000 10 MGGTIPHVSP 0.000

TABLE XV-V13 HLA-A11-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 10 FLPNGINGIK 0.400 9 TFLPNGINGI 0.003 2 SPKSLSETFL 0.002 8 ETFLPNGING 0.001 1 GSPKSLSETF 0.001 5 SLSETFLPNG 0.000 6 LSETFLPNGI 0.000 4 KSLSETFLPN 0.000 7 SETFLPNGIN 0.000 3 PKSLSETFLP 0.000

TABLE XV-V14 HLA-A11-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 3 LPLRLFTFWR 0.180 6 RLFTFWRGPV 0.024 8 FTFWRGPVVV 0.020 9 TFWRGPVVVA 0.004 2 NLPLRLFTFW 0.004 7 LFTFWRGPVV 0.002 1 ENLPLRLFTF 0.001 10 FWRGPVVVAI 0.000 4 PLRLFTFWRG 0.000 5 LRLFTFWRGP 0.000

TABLE XV-V21 HLA-A11-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 9 KTKHCMFSLI 0.030 2 SKLTQEQKTK 0.015 3 KLTQEQKTKH 0.012 5 TQEQKTKHCM 0.006 8 QKTKHCMFSL 0.001 6 QEQKTKHCMF 0.001 4 LTQEQKTKHC 0.001 7 EQKTKHCMFS 0.000 10 TKHCMFSLIS 0.000 1 LSKLTQEQKT 0.000

TABLE XV-V25 HLA-A11-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 2 ILFLPCISQK 0.800 4 FLPCISQKLK 0.200 5 LPCISQKLKR 0.080 8 ISQKLKRIKK 0.040 7 CISQKLKRIK 0.040 3 LFLPCISQKL 0.003 1 IILFLPCISQ 0.001 9 SQKLKRIKKG 0.000 10 QKLKRIKKGW 0.000 6 PCISQKLKRI 0.000

TABLE XVI-V8 HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 KSQFLEEGM 1.800 4 FLEEGMGGT 0.180 5 LEEGMGGTI 0.150 9 MGGTIPHVS 0.140 8 GMGGTIPHV 0.100 3 QFLEEGMGG 0.090 7 EGMGGTIPH 0.015 2 SQFLEEGMG 0.010 6 EEGMGGTIP 0.001

TABLE XVI-V13 HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 SPKSLSETF 2.400 9 FLPNGINGI 1.800 4 SLSETFLPN 0.144 6 SETFLPNGI 0.144 7 ETFLPNGIN 0.100 8 TFLPNGING 0.090 2 PKSLSETFL 0.040 3 KSLSETFLP 0.030 5 LSETFLPNG 0.015

TABLE XVI-V14 HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 NLPLRLFTF 3.000 8 TFWRGPVVV 0.500 6 LFTFWRGPV 0.500 2 LPLRLFTFW 0.216 7 FTFWRGPVV 0.100 9 FWRGPVVVA 0.100 5 RLFTFWRGP 0.020 4 LRLFTFWRG 0.002 3 PLRLFTFWR 0.001

TABLE XVI-V21 HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 8 KTKHCMFSL 8.000 6 EQKTKHCMF 2.000 4 TQEQKTKHC 0.150 9 TKHCMFSLI 0.120 5 QEQKTKHCM 0.075 2 KLTQEQKTK 0.020 1 SKLTQEQKT 0.020 3 LTQEQKTKH 0.020 7 QKTKHCMFS 0.010

TABLE XVI-V25 HLA-A24-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 3 FLPCISQKL 11.088 6 CISQKLKRI 1.000 2 LFLPCISQK 0.090 7 ISQKLKRIK 0.018 8 SQKLKRIKK 0.011 1 ILFLPCISQ 0.010 4 LPCISQKLK 0.010 9 QKLKRIKKG 0.002 5 PCISQKLKR 0.002

TABLE XVII-V8 HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 FLEEGMGGTI 1.800 4 QFLEEGMGGT 0.900 8 EGMGGTIPHV 0.150 9 GMGGTIPHVS 0.140 1 EKSQFLEEGM 0.060 2 KSQFLEEGMG 0.030 10 MGGTIPHVSP 0.010 3 SQFLEEGMGG 0.010 6 LEEGMGGTIP 0.002 7 EEGMGGTIPH 0.001

TABLE XVII-V13 HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 9 TFLPNGINGI 10.800 2 SPKSLSETFL 4.000 1 GSPKSLSETF 3.600 6 LSETFLPNGI 2.160 4 KSLSETFLPN 0.360 10 FLPNGINGIK 0.021 5 SLSETFLPNG 0.012 7 SETFLPNGIN 0.010 8 ETFLPNGING 0.010 3 PKSLSETFLP 0.000

TABLE XVII-V14 HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 1 ENLPLRLFTF 3.600 10 FWRGPVVVAI 1.400 7 LFTFWRGPVV 0.500 9 TFWRGPVVVA 0.500 2 NLPLRLFTFW 0.216 6 RLFTFWRGPV 0.200 8 FTFWRGPVVV 0.100 3 LPLRLFTFWR 0.015 5 LRLFTFWRGP 0.002 4 PLRLFTFWRG 0.001

TABLE XVII-V21 HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 9 KTKHCMFSLI 2.400 5 TQEQKTKHCM 0.750 8 QKTKHCMFSL 0.400 6 QEQKTKHCMF 0.300 4 LTQEQKTKHC 0.180 1 LSKLTQEQKT 0.132 7 EQKTKHCMFS 0.100 3 KLTQEQKTKH 0.022 10 TKHCMFSLIS 0.010 2 SKLTQEQKTK 0.002

TABLE XVII-V25 HLA-A24-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 3 LFLPCISQKL 66.528 6 PCISQKLKRI 0.150 10 QKLKRIKKGW 0.021 8 ISQKLKRIKK 0.017 4 FLPCISQKLK 0.015 1 IILFLPCISQ 0.015 7 CISQKLKRIK 0.012 9 SQKLKRIKKG 0.011 5 LPCISQKLKR 0.011 2 ILFLPCISQK 0.010

TABLE XVIII-V8 HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 KSQFLEEGM 1.000 8 GMGGTIPHV 0.200 7 EGMGGTIPH 0.030 4 FLEEGMGGT 0.030 9 MGGTIPHVS 0.020 5 LEEGMGGTI 0.012 2 SQFLEEGMG 0.010 6 EEGMGGTIP 0.001 3 QFLEEGMGG 0.001

TABLE XVIII-V13 HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 9 FLPNGINGI 0.400 1 SPKSLSETF 0.400 6 SETFLPNGI 0.040 2 PKSLSETFL 0.040 7 ETFLPNGIN 0.030 4 SLSETFLPN 0.020 3 KSLSETFLP 0.010 5 LSETFLPNG 0.003 8 TFLPNGING 0.001

TABLE XVIII-V14 HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 2 LPLRLFTFW 0.400 7 FTFWRGPVV 0.200 9 FWRGRVVVA 0.150 6 LFTFWRGPV 0.030 8 TFWRGPVVV 0.020 1 NLPLRLFTF 0.020 3 PLRLFTFWR 0.010 5 RLFTFWRGP 0.010 4 LRLFTFWRG 0.001

TABLE XVIII-V21 HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 8 KTKHCMFSL 4.000 5 QEQKTKHCM 0.100 9 TKHCMFSLI 0.040 4 TQEQKTKHC 0.030 6 EQKTKHCMF 0.020 3 LTQEQKTKH 0.010 1 SKLTQEQKT 0.010 2 KLTQEQKTK 0.010 7 QKTKHCMFS 0.002

TABLE XVIII-V25 HLA-B7-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 3 FLPCISQKL 4.000 6 CISQKLKRI 0.400 4 LPCISQKLK 0.200 8 SQKLKRIKK 0.015 1 ILFLPCISQ 0.015 7 ISQKLKRIK 0.010 9 QKLKRIKKG 0.001 2 LFLPCISQK 0.001 5 PCISQKLKR 0.001

TABLE XIX-V8 HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 8 EGMGGTIPHV 0.600 5 FLEEGMGGTI 0.120 1 EKSQFLEEGM 0.100 9 GMGGTIPHVS 0.020 10 MGGTIPHVSP 0.015 4 QFLEEGMGGT 0.010 3 SQFLEEGMGG 0.010 2 KSQFLEEGMG 0.010 7 EEGMGGTIPH 0.001 6 LEEGMGGTIP 0.000

TABLE XIX-V13 HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 2 SPKSLSETFL 80.000 6 LSETFLPNGI 0.120 9 TFLPNGINGI 0.040 1 GSPKSLSETF 0.020 4 KSLSETFLPN 0.020 10 FLPNGINGIK 0.010 5 SLSETFLPNG 0.010 8 ETFLPNGING 0.010 7 SETFLPNGIN 0.003 3 PKSLSETFLP 0.000

TABLE XIX-V14 HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 10 FWRGPVVVAI 0.400 6 RLFTFWRGPV 0.300 8 FTFWRGPVVV 0.200 3 LPLRLFTFWR 0.200 2 NLPLRLFTFW 0.020 7 LFTFWRGPVV 0.020 1 ENLPLRLFTF 0.020 9 TFWRGPVVVA 0.015 4 PLRLFTFWRG 0.010 5 LRLFTFWRGP 0.001

TABLE XIX-V21 HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 9 KTKHCMFSLI 0.400 8 QKTKHCMFSL 0.400 5 TQEQKTKHCM 0.300 1 LSKLTQEQKT 0.100 4 LTQEQKTKHC 0.100 7 EQKTKHCMFS 0.020 3 KLTQEQKTKH 0.010 10 TKHCMFSLIS 0.002 6 QEQKTKHCMF 0.002 2 SKLTQEQKTK 0.001

TABLE XIX-V25 HLA-B7-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 3 LFLPCISQKL 0.400 5 LPCISQKLKR 0.200 6 PCISQKLKRI 0.040 8 ISQKLKRIKK 0.015 1 IILFLPCISQ 0.015 7 CISQKLKRIK 0.010 4 FLPCISQKLK 0.010 9 SQKLKRIKKG 0.010 2 ILFLPCISQK 0.010 10 QKLKRIKKGW 0.002

TABLE XX-V8 HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 KSQFLEEGM 20.000 8 GMGGTIPHV 0.200 9 MGGTIPHVS 0.100 4 FLEEGMGGT 0.060 2 SQFLEEGMG 0.015 5 LEEGMGGTI 0.012 7 EGMGGTIPH 0.010 3 QFLEEGMGG 0.003 6 EEGMGGTIP 0.001

TABLE XX-V13 HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 1 SPKSLSETF 60.000 9 FLPNGINGI 0.400 4 SLSETFLPN 0.200 3 KSLSETFLP 0.150 7 ETFLPNGIN 0.100 6 SETFLPNGI 0.040 5 LSETFLPNG 0.015 2 PKSLSETFL 0.010 8 TFLPNGING 0.001

TABLE XX-V14 HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 2 LPLRLFTFW 10.000 1 NLPLRLFTF 1.000 7 FTFWRGPVV 0.200 9 FWRGPVVVA 0.030 6 LFTFWRGPV 0.020 5 RLFTFWRGP 0.020 8 TFWRGPVVV 0.020 3 PLRLFTFWR 0.003 4 LRLFTFWRG 0.001

TABLE XX-V21 HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 8 KTKHCMFSL 6.000 6 EQKTKHCMF 3.000 5 QEQKTKHCM 0.200 9 TKHCMFSLI 0.040 2 KLTQEQKTK 0.030 4 TQEQKTKHC 0.030 3 LTQEQKTKH 0.020 7 QKTKHCMFS 0.010 1 SKLTQEQKT 0.010

TABLE XX-V25 HLA-B3501-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Start Subsequence Score 3 FLPCISQKL 1.000 6 CISQKLKRI 0.400 4 LPCISQKLK 0.200 7 ISQKLKRIK 0.050 8 SQKLKRIKK 0.030 1 ILFLPCISQ 0.010 9 QKLKRIKKG 0.001 2 LFLPCISQK 0.001 5 PCISQKLKR 0.001

TABLE XXI-V8 HLA-B35-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 FLEEGMGGTI 0.240 8 EGMGGTIPHV 0.200 1 EKSQFLEEGM 0.200 2 KSQFLEEGMG 0.150 9 GMGGTIPHVS 0.100 4 QFLEEGMGGT 0.020 3 SQFLEEGMGG 0.015 10 MGGTIPHVSP 0.010 7 EEGMGGTIPH 0.001 6 LEEGMGGTIP 0.000

TABLE XXI-V13 HLA-B35-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 2 SPKSLSETFL 60.000 1 GSPKSLSETF 5.000 4 KSLSETFLPN 1.000 6 LSETFLPNGI 0.600 9 TFLPNGINGI 0.040 5 SLSETFLPNG 0.020 10 FLPNGINGIK 0.010 7 SETFLPNGIN 0.010 8 ETFLPNGING 0.010 3 PKSLSETFLP 0.000

TABLE XXI-V14 HLA-B35-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine. Start Subsequence Score 1 ENLPLRLFTF 1.000 2 NLPLRLFTFW 0.500 6 RLFTFWRGPV 0.400 8 FTFWRGPVVV 0.200 3 LPLRLFTFWR 0.200 10 FWRGPVVVAI 0.120 7 LFTFWRGPVV 0.020 9 TFWRGPVVVA 0.010 4 PLRLFTFWRG 0.003 5 LRLFTFWRGP 0.001

TABLE XXI-V21 HLA-B35-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine. Start Subsequence Score 9 KTKHCMFSLI 2.400 1 LSKLTQEQKT 1.500 5 TQEQKTKHCM 0.600 7 EQKTKHCMFS 0.300 4 LTQEQKTKHC 0.200 6 QEQKTKHCMF 0.100 8 QKTKHCMFSL 0.100 3 KLTQEQKTKH 0.020 10 TKHCMFSLIS 0.010 2 SKLTQEQKTK 0.002

TABLE XXI-V25 HLA-B35-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Start Subsequence Score 5 LPCISQKLKR 0.200 3 LFLPCISQKL 0.100 8 ISQKLKRIKK 0.050 10 QKLKRIKKGW 0.050 6 PCISQKLKRI 0.040 9 SQKLKRIKKG 0.030 4 FLPCISQKLK 0.010 7 CISQKLKRIK 0.010 2 ILFLPCISQK 0.010 1 IILFIPCISQ 0.010

TABLE XXII-V1 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 158 PKDASRQVY 27 419 FEEEYYRFY 27 405 ISTFHVLIY 26 221 SLATFFFLY 23 263 AITLLSLVY 23 392 SFIQSTLGY 23 276 LAAAYQLYY 22 280 YQLYYGTKY 21 244 QSDFYKIPI 19 101 LWDLRHLLV 18 189 PIDLGSLSS 18 198 AREIENLPL 18 231 FVRDVIHPY 18 240 ARNQQSDFY 18 275 LLAAAYQLY 18 311 FFAMVHVAY 18 90 FVAIHREHY 17 117 SNNMRINQY 17 327 RSERYLFLN 17 388 WREFSFIQS 17 427 YTPPNFVLA 17 443 ILDLLQLCR 17 444 LDLLQLCRY 17 46 TIRLIRCGY 16 66 ASEFFPHVV 16 124 QYPESNAEY 16 200 EIENLPLRL 16 330 RYLFLNMAY 16 352 EEVWRIEMY 16 272 LAGLLAAAY 15 323 LPMRRSERY 15 351 EEEVWRIEM 15 415 WKRAFEEEY 15 416 KRAFEEEYY 15 13 LSETCLPNG 14 38 SGDFAKSLT 14 98 YTSLWDLRH 14 178 VIELARQLN 14 406 STFHVLIYG 14 94 HREHYTSLW 13 135 SLFPDSLIV 13 137 FPDSLIVKG 13 251 PIEIVNKTL 13 396 STLGYVALL 13

TABLE XII-V2 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 23 LSLPSSWDY 23 36 PCPADFFLY 20 17 FTPFSCLSL 13 28 SWDYRCPPP 12

TABLE XXII-V5A HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 7 FTFWRGPVV 9 9 FWRGPVVVA 5

TABLE XXII-V5B HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 21 ELEFVFLLT 24 1 WREFSFIQI 17 17 QTELELEFV 16 13 FADTQTELE 15 19 ELELEFVFL 14

TABLE XXII-V6 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 34 FLEEGIGGT 14 28 GWEKSQFLE 12 35 LEEGIGGTI 12 29 WEKSQFLEE 11 41 GTIPHVSPE 11 1 VLPSIVILG 9 9 GKIILFLPC 9 19 SRKLKRIKK 9 2 LPSIVILGK 8 6 VILGKIILF 8 16 PCISRKLKR 8 7 ILGKIILFL 7 37 EGIGGTIPH 7 46 VSPERVTVM 7 3 PSIVILGKI 6 5 IVILGKIIL 6 12 ILFLPCISR 6

TABLE XXII-V7A HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 LSETFLPNG 14 4 SLSETFLPN 12 8 TFLPNGING 9 7 ETFLPNGIN 8 3 KSLSETFLP 6

TABLE XXII-V7B HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 AYQQSTLGY 22 9 STLGYVALL 13

TABLE XXII-V7C HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 59 WTEEAGATA 17 90 VTEDDEAQD 17 99 SIDPPESPD 17 167 KLETIILSK 17 32 LSEIVLPIE 16 51 STPPPPAMW 14 154 WSLGEFLGS 14 5 ILDLSVEVL 13 69 AQESGIRNK 13 9 SVEVLASPA 12 38 PIEWQQDRK 12 60 TEEAGATAE 12 66 TAEAQESGI 12 93 DDEAQDSID 12 104 ESPDRALKA 12 105 SPDRALKAA 12 123 HTNGVGPLW 12 130 LWEFLLRLL 12 96 AQDSIDPPE 11 102 PPESPDRAL 11 128 GPLWEFLLR 11 143 ASGTLSLAF 11 156 LGEFLGSGT 11 42 QQDRKIPPL 10 78 SSSSSQIPV 10 82 SQIPVVGVV 10 91 TEDDEAQDS 10 92 EDDEAQDSI 10 115 SWRNPVLPH 10 176 LTQEQKSKH 10 177 TQEQKSKHC 10 26 NILRGGLSE 9 50 LSTPPPPAM 9 79 SSSSQIPVV 9 131 WEFLLRLLK 9 2 SIVILDLSV 8 7 DLSVEVLAS 8 21 KCLGANILR 8 31 GLSEIVLPI 8 81 SSQIPVVGV 8 124 TNGVGPLWE 8 132 EFLLRLLKS 8 141 QAASGTLSL 8 162 SGTWMKLET 8 169 ETIILSKLT 8

TABLE XXII-V8 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 FLEEGMGGT 14 5 LEEGMGGTI 12 7 EGMGGTIPH 7

TABLE XXII-V13 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 LSETFLPNG 14 4 SLSETFLPN 12 8 TFLPNGING 9 7 ETFLPNGIN 8 3 KSLSETFLP 6

TABLE XXII-V14 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 7 FTFWRGPVV 9 9 FWRGPVVVA 5

TABLE XXII-V21 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 3 LTQEQKTKH 10 4 TQEQKTKHC 10 1 SKLTQEQKT 6 8 KTKHCMFSL 6 9 TKHCMFSLI 5

TABLE XXII-V25 HLA-A1-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 PCISQKLKR 10 8 SQKLKRIKK 9 1 ILFLPCISQ 6 2 LFLPCISQK 4 3 FLPCISQKL 4 7 ISQKLKRIK 4

TABLE XXIII-V1 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 365 IMSLGLLSL 29 271 YLAGLLAAA 28 433 VLALVLPSI 28 227 FLYSFVRDV 27 360 YISFGIMSL 27 396 STLGYVALL 27 17 CLPNGINGI 26 100 SLWDLRHLL 26 135 SLFPDSLIV 26 203 NLPLRLFTL 26 402 ALLISTFHV 26 436 LVLPSIVIL 26 128 SNAEYLASL 25 140 SLIVKGFNV 25 187 FIPIDLGSL 25 210 TLWRGPVVV 25 261 IVAITLLSL 25 403 LLISTFHVL 25 5 SMMGSPKSL 24 264 ITLLSLVYL 24 274 GLLAAAYQL 24 307 LLSFFFAMV 24 369 GLLSLLAVT 24 48 RLIRCGYHV 23 49 LIRCGYHVV 23 141 LIVKGFNVV 23 313 AMVHVAYSL 23 374 LAVTSIPSV 23 393 FIQSTLGYV 23 441 IVILDLLQL 23 106 HLLVGKILI 22 180 ELARQLNFI 22 254 IVNKTLPIV 22 258 TLPIVAITL 22 262 VAITLLSLV 22 265 TLLSLVYLA 22 267 LSLVYLAGL 22 268 SLVYLAGLL 22 333 FLNMAYQQV 22 378 SlPSVSNAL 22 404 LISTFHVLI 21 435 ALVLPSIVI 21 107 LLVGKILID 20 108 LVGKILIDV 20 112 ILIDVSNNM 20 173 QARQQVIEL 20 184 QLNFIPIDL 20 368 LGLLSLLAV 20 65 FASEFFPHV 19 83 LTKTNIIFV 19 133 LASLFPDSL 19 177 QVIELARQL 19 257 KTLPIVAIT 19 306 GLLSFFFAM 19 366 MSLGLLSLL 19 434 LALVLPSIV 19 27 DARKVTVGV 18 196 SSAREIENL 18 209 FTLWRGPVV 18 259 LPIVAITLL 18 367 SLGLLSLLA 18 371 LSLLAVTSI 18 397 TLGYVALLI 18 41 FAKSLTIRL 17 81 DALTKTNII 17 85 KTNIIFVAI 17 103 DLRHLLVGK 17 104 LRHLLVGKI 17 153 ALQLGPKDA 17 155 QLGPKDASR 17 212 WRGPVVVAI 17 250 IPIEIVNKT 17 253 EIVNKTLPI 17 363 FGIMSLGLL 17 370 LLSLLAVTS 17 410 VLIYGWKRA 17 428 TPPNFVLAL 17 438 LPSIVILDL 17 442 VILDLLQLC 17 25 IKDARKVTV 16 68 EFFPHVVDV 16 88 IIFVAIHRE 16 93 IHREHYTSL 16 99 TSLWDLRHL 16 132 YLASLFPDS 16 148 VVSAWALQL 16 171 NIQARQQVI 16 190 IDLGSLSSA 16 200 EIENLPLRL 16 372 SLLAVTSIP 16 12 SLSETCLPN 15 44 SLTIRLIRC 15 50 IRCGYHVVI 15 111 KILIDVSNN 15 211 LWRGPVVVA 15 217 VVAISLATF 15 221 SLATFFFLY 15 247 FYKIPIEIV 15 249 KIPIEIVNK 15 251 PIEIVNKTL 15 256 NKTLPIVAI 15 270 VYLAGLLAA 15 299 LQCRKQLGL 15 324 PMRRSERYL 15 331 YLFLNMAYQ 15 335 NMAYQQVHA 15 385 ALNWREFSF 15 400 YVALLISTF 15 437 VLPSIVILD 15 23 NGIKDARKV 14 37 GSGDFAKSL 14 39 GDFAKSLTI 14 42 AKSLTIRLI 14 164 QVYICSNNI 14 166 YICSNNIQA 14 220 ISLATFFFL 14 223 ATFFFLYSF 14 266 LLSLVYLAG 14 275 LLAAAYQLY 14 278 AAYQLYYGT 14 300 QCRKQLGLL 14 309 SFFFAMVHV 14 362 SFGIMSLGL 14 373 LLAVTSIPS 14 395 QSTLGYVAL 14 411 LIYGWKRAF 14 427 YTPPNFVLA 14 443 ILDLLQLCR 14

TABLE XXIII-V2 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 GLQALSLSL 25 1 SGSPGLQAL 21 8 ALSLSLSSG 18 17 FTPFSCLSL 17 10 SLSLSSGFT 16 3 SPGLQALSL 15 12 SLSSGFTPF 14 15 SGFTPFSCL 14 24 SLPSSWDYR 12

TABLE XXIII-V5A HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 7 FTFWRGPVV 17 1 NLPLRLFTF 16 8 TFWRGPVVV 15 9 FWRGPVVVA 14 5 RLFTFWRGP 13 3 PLRLFTFWR 10 6 LFTFWRGPV 10

TABLE XXIII-V5B HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 20 LELEFVFLL 21 22 LEFVFLLTL 21 24 FVFLLTLLL 20 19 ELELEFVFL 18 12 SFADTQTEL 17 17 QTELELEFV 17 8 QIFCSEADT 15 6 FIQIFCSFA 14 14 ADTQTELEL 14 23 EFVFLLTLL 11 21 ELEFVFLLT 10

TABLE XXIII-V6 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 7 ILGKIILFL 27 38 GIGGTIPHV 26 10 KIILFLPCI 25 14 FLPCISRKL 23 34 FLEEGIGGT 23 5 IVILGKIIL 20 17 CISRKLKRI 20 45 HVSPERVTV 20 4 SIVILGKII 18 6 VILGKIILF 18 12 ILFLPCISR 16 1 VLPSIVILG 15 27 KGWEKSQFL 15 3 PSIVILGKI 13 35 LEEGIGGTI 13 41 GTIPHVSPE 13

TABLE XXIII-V7A HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 FLPNGINGI 27 4 SLSETFLPN 15

TABLE XXIII-V7B HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 STLGYVALL 27 3 NMAYQQSTL 21 6 YQQSTLGYV 16 8 QSTLGYVAL 14

TABLE XXIII-V7C HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 27 ILRGGLSEI 30 4 VILDLSVEV 27 4 VILDLSVEV 27 5 ILDLSVEVL 26 31 GLSEIVLPI 26 129 PLWEFLLRL 26 148 SLAFTSWSL 25 2 SIVILDLSV 24 141 QAASGTLSL 23 155 SLGEFLGSG 21 163 GTWMKLETI 21 81 SSQIPVVGV 20 82 SQIPVVGVV 20 119 PVLPHTNGV 19 133 FLLRLLKSQ 19 165 WMKLETIIL 19 24 GANILRGGL 18 57 AMWTEEAGA 18 112 AANSWRNPV 18 126 GVGPLWEFL 18 12 VLASPAAAW 17 79 SSSSQIPVV 17 134 LLRLLKSQA 17 167 KLETIILSK 17 168 LETIILSKL 17 171 IILSKLTQE 17 172 ILSKLTQEQ 17 42 QQDRKIPPL 16 142 AASGTLSLA 16 160 LGSGTWMKL 16 7 DLSVEVLAS 15 17 AAAWKCLGA 15 22 CLGANILRG 15 26 NILRGGLSE 15 28 LRGGLSEIV 15 130 LWEFLLRLL 15 136 RLLKSQAAS 15 137 LLKSQAASG 15 159 FLGSGTWMK 15 185 CMFSLISGS 15 83 QIPVVGVVT 14

TABLE XXIII-V8 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 GMGGTIPHV 26 4 FLEEGMGGTI 19 5 LEEGMGGGTI 13

TABLE XXIII-V13 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 FLPNGINGI 27 4 SLSETFLPN 15

TABLE XXIII-V14 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 7 FTFWRGPVV 17 1 NLPLRLFTF 16 8 TFWRGPVVV 15 9 FWRGPVVVA 14 5 RLFTFWRGP 13 3 PLRLETFWR 10 6 LFTFWRGPV 10

TABLE XXIII-V21 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 KTKHCMFSL 16 2 KLTQEQKTK 11 1 SKLTQEQKT 10 3 LTQEQKTKH 10 9 TKHCMFSLI 8

TABLE XXIII-V25 HLA-A0201-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 3 FLPCISQKL 23 6 CISQKLKRI 20 1 ILFLPCISQ 16

TABLE XXIV-V1 HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V2 HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V5A HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V5B HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V6 HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V7A HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V7B HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V7C HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V8 HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V13 HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V14 HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V21 HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXIV-V25 HLA-A0203-9mers-98P4B6 Pos 123456789 score NoResultsFound.

TABLE XXV-V1 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 103 DLRHLLVGK 27 56 VVIGSRNPK 26 249 KIPIEIVNK 26 3 SISMMGSPK 25 155 QLGPKDASR 25 263 AITLLSLVY 25 210 TLWRGPVVV 24 48 RLIRCGYHV 23 142 IVKGFNVVS 23 217 VVAISLATF 23 400 YVALLISTF 23 177 QVIELARQL 22 205 PLRLFTLWR 22 281 QLYYGTKYR 22 370 LLSLLAVTS 22 441 IVILDLLQL 22 35 VIGSGDFAK 21 77 THHEDALTK 21 148 VVSAWALQL 21 231 FVRDVIHPY 21 269 LVYLAGLLA 21 375 AVTSIPSVS 21 385 ALNWREFSF 21 274 GLLAAAYQL 20 322 CLPMRRSER 20 409 HVLIYGWKR 20 443 ILDLLQLCR 20 46 TIRLIRCGY 19 87 NIIFVAIHR 19 90 FVAIHREHY 19 258 TLPIVAITL 19 261 IVAITLLSL 19 275 LLAAAYQLY 19 279 AYQLYYGTK 19 369 GLLSLLAVT 19 372 SLLAVTSIP 19 411 LIYGWKRAF 19 436 LVLPSIVIL 19 34 GVIGSGDFA 18 92 AIHREHYTS 18 140 SLIVKGFNV 18 191 DLGSLSSAR 18 221 SLATFFFLY 18 435 ALVLPSIVI 18 22 INGIKDARK 17 49 LIRCGYHVV 17 82 ALTKTNIIF 17 111 KILIDVSNN 17 112 ILIDVSNNM 17 315 SLFPDSLIV 17 153 ALQLGPKDA 17 164 QVYICSNNI 17 203 NLPLRLFTL 17 271 YLAGLLAAA 17 304 QLGLLSFFF 17 381 SVSNALNWR 17 397 TLGYVALLI 17 403 LLISTFHVL 17 432 FVLALVLPS 17 32 TVGVIGSGD 16 107 LLVGKILID 16 151 AWALQLGPK 16 171 NIQARQQVI 16 189 PIDLGSLSS 16 216 VVVAISLAT 16 219 AISLATFFF 16 234 DVIHPYARN 16 266 LLSLVYLAG 16 302 RKQLGLLSF 16 402 ALLISTFHV 16 12 SLSETCLPN 15 21 GINGIKDAR 15 24 GIKDARKVT 15 30 KVTVGVIGS 15 121 RINQYPESN 15 136 LFPDSLIVK 15 179 IELARQLNF 15 268 SLVYLAGLL 15 356 RIEMYISFG 15 367 SLGLLSLLA 15 410 VLIYGWKRA 15 433 VLALVLPSI 15 25 IKDARKVTV 14 44 SLTIRLIRC 14 57 VIGSRNPKF 14 61 RNPKFASEF 14 106 HLLVGKILI 14 141 LIVKGFNVV 14 180 ELARQLNFI 14 207 RLFTLWRGP 14 227 FLYSFVRDV 14 235 VIHPYARNQ 14 241 RNQQSDFYK 14 251 PIEIVNKTL 14 272 LAGLLAAAY 14 294 WLETWLQCR 14 303 KQLGLLSFF 14 307 LLSFFFAMV 14 330 RYLFLNMAY 14 331 YLFLNMAYQ 14 340 QVHANIENS 14 353 EVWRIEMYI 14 364 GIMSLGLLS 14 17 CLPNGINGI 13 18 LPNGINGIK 13 26 KDARKVTVG 13 43 KSLTIRLIR 13 55 HVVIGSRNP 13 70 FPHVVDVTH 13 100 SLWDLRHLL 13 113 LIDVSNNMR 13 147 NYVSAWALQ 13 158 PKDASRQVY 13 184 QLNFIPIDL 13 200 EIENLPLRL 13 211 LWRGPVVVA 13 215 PVVVAISLA 13 253 EIVNKTLPI 13 260 PIVAITLLS 13 306 GLLSFFFAM 13 311 FFAMVHVAY 13 314 MVHVAYSLC 13 333 FLNMAYQQV 13 360 YISFGIMSL 13 392 SFIQSTLGY 13 408 FHVLIYGWK 13 440 SIVILDLLQ 13

TABLE XXV-V2-HLA HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 ALSLSLSSG 19 12 SLSSGFTPF 18 5 GLQALSLSL 17 22 CLSLPSSWD 15 24 SLPSSWDYR 15 10 SLSLSSGFT 13 23 LSLPSSWDY 11 33 CPPPCPADF 11 3 SPGLQALSL 10 7 QALSLSLSS 9 9 LSLSLSSGF 9 11 LSLSSGFTP 9 21 SCLSLPSSW 9 37 CPADFFLYF 9

TABLE XXV-V5A HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 NLPLRLFTF 21 3 PLRLFTFWR 19 5 RLFTFWRGP 14 8 TFWRGPVVV 14 9 FWRGPVVVA 13

TABLE XXV-V5B HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 19 ELELEFVFL 15 21 ELEFVFLLT 14 24 FVELLTLLL 14 8 QIFCSFADT 13 6 FIQIFCSFA 12 18 TELELEFVF 11 5 SFIQIFCSF 10 9 IFCSFADTQ 9 2 REFSFIQIF 8 16 TQTELELEF 8 22 LEFVFLLTL 7

TABLE XXV-V6 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 45 HVSPERVTV 22 23 KRIKKGWEK 20 12 ILFLPCISR 19 5 IVILGKIIL 18 13 LFLPCISRK 18 6 VILGKIILF 17 21 KLKRIKKGW 17 2 LPSIVILGK 15 7 ILGKIILFL 15 10 KIILFLPCI 15 18 ISRKLKRIK 15 19 SRKLKRIKK 15 24 RIKKGWEKS 15 34 FLEEGIGGT 14 4 SIVILGKII 13 11 IILFLPCIS 13 26 KKGWEKSQF 13 42 TIPHVSPER 13 15 LPCISRKLK 12 16 PCISRKLKR 12 17 PCISRKLKR 12 37 EGIGGTIPH 11 1 VLPSIVILG 10 14 FLPCISRKL 10 35 LEEGIGGTI 10 38 GIGGTIPHV 10

TABLE XXV-V7A HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 SLSETFLPN 15 9 FLPNGINGI 13 1 SPKSLSETF 10 8 TFLPNGING 8

TABLE XXV-V7B HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 FLNMAYQQS 13 5 AYQQSTLGY 12 8 QSTLGYVAL 10 7 QQSTLGYVA 9 3 NMAYQQSTL 8 9 STLGYVALL 8 4 MAYQQSTLG

TABLE XXV-V7C HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 167 KLETIILSK 28 175 KLTQEQKSK 25 109 ALKAANSWR 24 3 IVILDLSVE 23 26 NILRGGLSE 23 159 FLGSGTWMK 23 27 ILRGGLSEI 22 83 QIPVVGVVT 22 13 LASPAAAWK 20 35 IVLPIEWQQ 20 134 LLRLLKSQA 20 136 RLLKSQAAS 20 11 EVLASPAAA 19 137 LLKSQAASG 19 170 TIILSKLTQ 19 12 VLASPAAAW 18 38 PIEWQQDRK 18 73 GIRNKSSSS 18 5 ILDLSVEVL 17 9 SVEVLASPA 17 45 RKIPPLSTP 17 103 PESPDRALK 17 133 FLLRLLKSQ 17 171 IILSKLTQE 17 2 SIVILDLSV 15 4 VILDLSVEV 15 22 CLGANILRG 15 46 KIPPLSTPP 15 69 AQESGIRNK 15 99 SIDPPESPD 15 119 PVLPHTNGV 15 120 VLPHTNGVG 15 131 WEFLLRLLK 15 155 SLGEFLGSG 15 173 LSKLTQEQK 15 7 DLSVEVLAS 14 31 GLSEIVLPI 14 36 VLPIEWQQD 14 85 PVVGVVTED 14 129 PLWEFLLRL 14 146 TLSLAFTSW 14 148 SLAFTSWSL 14 25 ANILRGGLS 13 82 SQIPVVGVV 13 126 GVGPLWEFL 13

TABLE XXV-V8 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 FLEEGMGGT 14 5 LEEGMGGTI 10 3 QFLEEGMGG 9 7 EGMGGTIPH 8 6 EEGMGGTIP 6

TABLE XXV-V13 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 SLSETFLPN 15 9 FLPNGINGI 13 1 SPKSLSETF 10 8 TFLPNGING 8

TABLE XXV-V14 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 NLPLRLFTF 21 3 PLRLFTFWR 19 5 RLFTFWRGP 14 8 TFWRGPVVV 14 9 FWRGPVVVA 13

TABLE XXV-V21 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 KLTQEQKTK 27

TABLE XXV-V25 HLA-A3-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 LFLPCISQK 21 1 ILFLPCISQ 15 8 SQKLKRIKK 15 7 ISQKLKRIK 12 4 LPCISQKLK 11 3 FLPCISQKL 10 5 PCISQKLKR 10

TABLE XXVI-V1 HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 352 EEVWRIEMY 29 75 DVTHHEDAL 28 441 IVILDLLQL 28 177 QVIELARQL 26 223 ATFFFLYSF 25 231 FVRDVIHPY 25 400 YVALLISTF 25 200 EIENLPLRL 24 261 IVAITLLSL 24 217 VVAISLATF 23 436 LVLPSIVIL 23 96 EHYTSLWDL 22 234 DVIHPYARN 22 353 EVWRIEMYI 22 390 EFSFIQSTL 22 396 STLGYVALL 21 90 FVAIHREHY 20 148 VVSAWALQL 20 253 EIVNKTLPI 20 264 ITLLSLVYL 20 15 ETCLPNGIN 19 68 EFFPHVVDV 19 115 DVSNNMRIN 19 215 PVVVAISLA 19 296 ETWLQCRKQ 19 31 VTVGVIGSG 18 187 FIPIDLGSL 18 216 VVVAISLAT 18 406 STFHVLIYG 18 439 PSIVILDLL 18 2 ESISMMGSP 17 45 LTIRLIRCG 17 46 TIRLIRCGY 17 108 LVGKILIDV 17 263 AITLLSLVY 17 360 YISFGIMSL 17 363 FGIMSLGLL 17 30 KVTVGVIGS 16 117 SNNMRINQY 16 128 SNAEYLASL 16 259 LPIVAITLL 16 355 WRIEMYISF 16 392 SFIQSTLGY 16 405 ISTFHVLIY 16 432 FVLALVLPS 16 32 TVGVIGSGD 15 34 GVIGSGDFA 15 72 HVVDVTHHE 15 147 NVVSAWALQ 15 257 KTLPIVAIT 15 268 SLVYLAGLL 15 329 ERYLFLNMA 15 340 QVHANIENS 15 375 AVTSIPSVS 15 378 SIPSVSNAL 15 381 SVSNALNWR 15 428 TPPNFVLAL 15 55 HVVIGSRNP 14 56 VVIGSRNPK 14 57 VIGSRNPKF 14 83 LTKTNIIFV 14 131 EYLASLFPD 14 138 PDSLIVKGF 14 180 ELARQLNFI 14 214 GPVVVAISL 14 218 VAISLATFF 14 254 IVNKTLPIV 14 302 RKQLGLLSF 14 303 KQLGLLSFF 14 316 HVAYSLCLP 14 365 IMSLGLLSL 14 366 MSLGLLSLL 14 430 PNFVLALVL 14 444 LDLLQLCRY 14

TABLE XXVI-V2 HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 17 FTPFSCLSL 18 1 SGSPGLQAL 15 15 SGFTPFSCL 14 3 SPGLQALSL 11 5 GLQALSLSL 11 9 LSLSLSSGF 11 18 TPFSCLSLP 11 23 LSLPSSWDY 11 12 SLSSGFTPF 10 36 PCPADFFLY 10 37 CPADFFLYF 10 33 CPPPCPADF 9 35 PPCPADFFL 9 30 DYRCPPPCP 8 34 PPPCPADFF 8

TABLE XXVI-V5A HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 NLPLRLFTF 13 7 FTFWRGPVV 13

TABLE XXVI-V5B HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 23 EFVFLLTLL 27 24 FVFLLTLLL 24 15 DTQTELELE 20 T9 ELELEFVFL 18 22 LEFVFLLTL 18 2 REFSFIQIF 17 5 SFIQIFCSF 16 16 TQTELELEF 14 20 LELEFVFLL 14 3 EFSFIQIFC 13

TABLE XXVI-V6 HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 IVILGKIIL 23 6 VILGKIILF 18 41 GTIPHVSPE 18 7 ILGKIILFL 15 37 EGIGGTIPH 15 30 EKSQFLEEG 14 3 PSIVILGKI 12 10 KIILFLPCI 12 45 HVSPERVTV 12 4 SIVILGKII 11 14 FLPCISRKL 11 27 KGWEKSQFL 11 36 EEGIGGTIP 11

TABLE XXVI-V7A HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 7 ETFLPNGIN 23 1 SPKSLSETF 12

TABLE XXVI-V7B HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 STLGYVALL 21 5 AVQQSTLGY 11 3 NMAYQQSTL 10 8 QSTLGYVAL 10

TABLE XXVI-V7C HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 169 ETIILSKLT 23 34 EIVLPIEWQ 22 11 EVLASPAAA 21 151 FTSWSLGEF 21 179 EQKSKHCMF 21 126 GVGPLWEFL 20 3 IVILDLSVE 19 85 PVVGVVTED 18 168 LETIILSKL 17 125 NGVGPLWEF 16 132 EFLLRLLKS 16 95 EAQDSIDPP 15 129 PLWEFLLRL 15 7 DLSVEVLAS 14 35 IVLPIEWQQ 14 68 EAQESGIRN 14 88 GVVTEDDEA 14 89 VVTEDDEAQ 14 98 DSIDPPESP 14 122 PHTNGVGPL 14 163 GTWMKLETI 14 9 SVEVLASPA 13 42 QQDRKIPPL 13 92 EDDEAQDSI 13 104 ESPDRALKA 13 130 LWEFLLRLL 13 2 SIVILDLSV 12 5 ILDLSVEVL 12 59 WTEEAGATA 12 152 TSWSLGEFL 12 176 LTQEQKSKH 12 8 LSVEVLASP 11 45 RKIPPLSTP 11 51 STPPPPAMW 11 62 EAGATAEAQ 11 65 ATAEAQESG 11 71 ESGIRNKSS 11 82 SQIPVVGVV 11 119 PVLPHTNGV 11 141 QAASGTLSL 11 143 ASGTLSLAF 11 145 GTLSLAFTS 11 158 EFLGSGTWM 11 170 TIILSKLTQ 11 171 IILSKLTQE 11 185 CMFSLISGS 11

TABLE XXVI-V8 HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 6 EEGMGGTIP 11 7 EGMGGTIPH 11 2 SQFLEEGMG 7

TABLE XXVI-V13 HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 7 ETFLPNGIN 23 1 SPKSLSETF 12

TABLE XXVI-V14 HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 NLPLRLFTF 13 7 FTFWRGPVV 13

TABLE XXVI-V21 HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 6 EQKTKHCMF 20 8 KTKHCMFSL 17 3 LTQEQKTKH 11

TABLE XXVI-V25 HLA-A26-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 3 FLPCISQKL 11 6 CISQKLKRI 9 2 LFLPCISQK 7 5 PCISQKLKR 7 1 ILFLPCISQ 6 9 QKLKRIKKG 5

TABLE XXVII-V1 HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 428 TPPNFVLAL 24 438 LPSIVILDL 24 259 LPIVAITLL 21 291 FPPWLETWL 21 125 YPESNAEYL 20 214 GPVVVAISL 20 250 IPIEIVNKT 18 62 NPKFASEFF 17 211 LWRGPVVVA 17 429 PPNFVLALV 17 157 GPKDASRQV 16 326 RRSERYLFL 16 148 VVSAWALQL 15 198 AREIENLPL 15 365 IMSLGLLSL 15 426 FYTPPNFVL 15 93 IHREHYTSL 14 220 ISLATFFFL 14 261 IVAITLLSL 14 287 KYRRFPPWL 14 379 IPSVSNALN 14 396 STLGYVALL 14 5 SMMGSPKSL 13 10 PKSLSETCL 13 137 FPDSLIVKG 13 173 QARQQVIEL 13 200 EIENLPLRL 13 264 ITLLSLVYL 13 289 RRFPPWLET 13 300 QCRKQLGLL 13 315 VHVAYSLCL 13 362 SFGIMSLGL 13 390 EFSFIQSTL 13 395 QSTLGYVAL 13 430 PNFVLALVL 13 436 LVLPSIVIL 13 441 IVILDLLQL 13 18 LPNGINGIK 12 27 DARKVTVGV 12 50 IRCGYHVVI 12 70 FPHVVDVTH 12 105 RHLLVGKIL 12 128 SNAEYLASL 12 133 LASLFPDSL 12 188 IPIDLGSLS 12 202 ENLPLRLFT 12 204 LPLRLFTLW 12 212 WRGPVVVAI 12 219 AISLATFFF 12 256 NKTLPIVAI 12 299 LQCRKQLGL 12 313 AMVHVAYSL 12 324 PMRRSERYL 12 360 YISFGIMSL 12 366 MSLGLLSLL 12 403 LLISTFHVL 12 435 ALVLPSIVI 12 25 IKDARKVTV 11 37 GSGDFAKSL 11 41 FAKSLTIRL 11 68 EFFPHVVDV 11 75 DVTHHEDAL 11 85 KTNIIFVAI 11 96 EHYTSLWDL 11 100 SLWDLRHLL 11 134 ASLFPDSLI 11 146 FNVVSAWAL 11 196 SSAREIENL 11 237 HPYARNQQS 11 253 EIVNKTLPI 11 267 LSLVYLAGL 11 271 YLAGLLAAA 11 274 GLLAAAYQL 11 292 PPWLETWLQ 11 297 TWLQCRKQL 11 323 LPMRRSERY 11 328 SERYLFLNM 11 378 SIPSVSNAL 11 394 IQSTLGYVA 11 425 RFYTPPNFV 11

TABLE XXVII-V2 HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 3 SPGLQALSL 23 35 PPCPADFFL 22 34 PPPCPADFF 20 37 CPADFFLYF 20 33 CPPPCPADF 18 1 SGSPGLQAL 14 15 SGFTPFSCL 14 5 GLQALSLSL 13 17 FTPFSCLSL 12 25 LPSSWDYRC 12 12 SLSSGFTPF 11 31 YRCPPPCPA 11

TABLE XXVII-V5A HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 FWRGPVVVA 17 2 LPLRLFTFW 13 7 FTFWRGPVV 9 8 TFWRGPVVV 9 6 LFTFWRGPV 8

TABLE XXVII-V5B HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 19 ELELEFVFL 15 14 ADTQTELEL 14 24 FVFLLTLLL 13 12 SFADTQTEL 12 22 LEFVFLLTL 12 23 EFVFLLTLL 12 20 LELEFVFLL 11 21 ELEFVFLLT 10 10 FCSFADTQT 9 8 QIFCSFADT 8 16 TQTELELEF 8 1 WREFSFIQI 7 2 REFSFIQIF 7 5 SFIQIFCSF 7 6 FIQIFCSFA 7 17 QTELELEFV 7 18 TELELEFVF 7

TABLE XXVII-V6 HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 43 IPHVSPERV 17 7 ILGKIILFL 16 2 LPSIVILGK 14 27 KGWEKSQFL 12 45 HVSPERVTV 12 5 IVILGKIIL 11 15 LPCISRKLK 11 14 FLPCISRKL 10 38 GIGGTIPHV 10 44 PHVSPERVT 10 35 LEEGIGGTI 9 46 VSPERVTVM 9 6 VILGKIILF 8 10 KIILFLPCI 8 17 CISRKLKRI 8 26 KKGWEKSQF 8

TABLE XXVII-V7 HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 SPKSLSETF 16 2 PKSLSETFL 14

TABLE XXVII-V7B HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 STLGYVALL 14 8 QSTLGYVAL 13 3 NMAYQQSTL 11 7 QQSTLGYVA 10 2 LNMAYQQST 8 6 YQQSTLGYV 6

TABLE XXVII-V7C HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 102 PPESPDRAL 24 15 SPAAAWKCL 22 52 TPPPPAMWT 20 55 PPAMWTEEA 18 105 SPDRALKAA 18 101 DPPESPDRA 16 113 ANSWRNPVL 16 5 ILDLSVEVL 14 47 IPPLSTPPP 14 84 IPVVGVVTE 14 118 NPVLPHTNG 14 141 QAASGTLSL 14 160 LGSGTWMKL 14 29 RGGLSEIVL 13 42 QQDRKIPPL 13 49 PLSTPPPPA 13 121 LPHTNGVGP 13 126 GVGPLWEFL 13 128 GPLWEFLLR 13 31 GLSEIVLPI 12 48 PPLSTPPPP 12 50 LSTPPPPAM 12 54 PPPAMWTEE 12 61 EEAGATAEA 12 81 SSQIPVVGV 12 122 PHTNGVGPL 12 129 PLWEFLLRL 12 139 KSQAASGTL 12 142 AASGTLSLA 12 143 ASGTLSLAF 12 152 TSWSLGEFL 12 17 AAAWKCLGA 11 24 GANILRGGL 11 27 ILRGGLSEI 11 44 DRKIPPLST 11 53 PPPPAMWTE 11 125 NGVGPLWEF 11 148 SLAFTSWSL 11 158 EFLGSGTWM 11 165 WMKLETIIL 11 181 KSKHCMFSL 11

TABLE XXVII-V8 HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 GMGGTIPHV 10 5 LEEGMGGTI 9 1 KSQFLEEGM 7 4 FLEEGMGGT 6 7 EGMGGTIPH 6 6 EEGMGGTIP 4

TABLE XXVII-V13 HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 SPKSLSETF 16 2 PKSLSETFL 14

TABLE XXVII-V14 HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 FWRGPVVVA 17 2 LPLRLFTFW 13 7 FTFWRGPVV 9 8 TFWRGPVVV 9 6 LFTFWRGPV 8

TABLE XXVII-V21 HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 KTKHCMFSL 11 5 QEQKTKHCM 7 6 EQKTKHCMF 7 9 TKHCMFSLI 7 1 SKLTQEQKT 6

TABLE XXVII-V25 HLA-B0702-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 3 FLPCISQKL 10 4 LPCISQKLK 10 6 CISQKLKRI 8 1 ILFLPCISQ 4

TABLE XXVIII-V1 HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 41 FAKSLTIRL 25 203 NLPLRLFTL 25 62 NPKFASEFF 22 173 QARQQVIEL 22 253 EIVNKTLPI 22 57 VIGSRNPKF 20 81 DALTKTNII 20 285 GTKYRRFPP 20 299 LQCRKQLGL 20 326 RRSERYLFL 20 385 ALNWREFSF 20 93 IHREHYTSL 19 140 SLIVKGFNV 19 268 SLVYLAGLL 19 9 SPKSLSETC 18 28 ARKVTVGVI 18 100 SLWDLRHLL 18 171 NIQARQQVI 18 214 GPVVVAISL 18 259 LPIVAITLL 18 428 TPPNFVLAL 18 39 GDFAKSLTI 17 107 LLVGKILID 17 157 GPKDASRQV 17 274 GLLAAAYQL 17 291 FPPWLETWL 17 378 SIPSVSNAL 17 438 LPSIVILDL 17 24 GIKDARKVT 16 44 SLTIRLIRC 16 125 YPESNAEYL 16 155 QLGPKDASR 16 184 QLNFIPIDL 16 200 EIENLPLRL 16 237 HPYARNQQS 16 239 YARNQQSDF 16 251 PIEIVNKTL 16 258 TLPIVAITL 16 283 YYGTKYRRF 16 287 KYRRFPPWL 16 300 QCRKQLGLL 16 324 PMRRSERYL 16 403 LLISTFHVL 16 133 LASLFPDSL 15 159 KDASRQVYI 15 179 IELARQLNF 15 187 FIPIDLGSL 15 322 CLPMRRSER 15 360 YISFGIMSL 15 106 HLLVGKILI 14 128 SNAEYLASL 14 180 ELARQLNFI 14 197 SAREIENLP 14 245 SDFYKIPIE 14 298 WLQCRKQLG 14 323 LPMRRSERY 14 433 VLALVLPSI 14 5 SMMGSPKSL 13 17 CLPNGINGI 13 82 ALTKTNIIF 13 91 VAIHREHYT 13 103 DLRHLLVGK 13 142 IVKGFNVVS 13 146 FNVVSAWAL 13 196 SSAREIENL 13 205 PLRLFTLWR 13 264 ITLLSLVYL 13 304 QLGLLSFFF 13 395 QSTLGYVAL 13 396 STLGYVALL 13 397 TLGYVALLI 13 435 ALVLPSIVI 13 37 GSGDFAKSL 12 60 SRNPKFASE 12 96 SHYTSLWDL 12 105 HRLLVGKIL 12 109 VGKILIDVS 12 177 QVIELARQL 12 247 FYKIPIEIV 12 325 MRRSERYLF 12 362 SFGIMSLGL 12 365 IMSLGLLSL 12 390 EFSFIQSTL 12 414 GWKRAFEEE 12 426 FYTPPNFVL 12 436 LVLPSIVIL 12 441 IVILDLLQL 12

TABLE XXVIII-V2 HLA-B08-9mers-98P4B6 Each pept1de is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 3 SPGLQALSL 19 5 GLQALSLSL 17 35 PPCPADFFL 16 12 SLSSGFTPF 14 1 SGSPGLQAL 13 15 SGFTPFSCL 12 33 CPPPCPADF 12 34 PPPCPADFF 12 37 CPADFFLYF 12 17 FTPFSCLSL 11 28 SWDYRCPPP 11 10 SLSLSSGFT 9

TABLE XXIX-V5A HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 NLPLRLFTF 21 3 PLRLFTFWR 13

TABLE XXIX-V5B HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 19 ELELEFVFL 20 12 SFADTQTEL 13 20 LELEFVFLL 13 23 EFVFLLTLL 12 24 FVFLLTLLL 12 14 ADTQTELEL 11 22 LEFVFLLTL 11 16 TQTELELEF 9

TABLE XXIX-V6 HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 19 SRKLKRIKK 23 6 VILGKIILF 22 27 KGWEKSQFL 22 17 CISRKLKRI 21 7 ILGKIILFL 18 14 FLPCISRKL 17 21 KLKRIKKGW 17 22 LKRIKKGWE 16 24 RIKKGWEKS 14 4 SIVILGKII 13 5 IVILGKIIL 12 25 IKKGWEKSQ 12 46 VSPERVTVM 12 10 KIILFLPCI 11 23 KRIKKGWEK 11 29 WEKSQFLEE 11

TABLE XXIX-V7A HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 SPKSLSETF 24 9 FLPNGINGI 14 2 PKSLSETFL 11

TABLE XXIX-V7B HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 QSTLGYVAL 13 9 STLGYVALL 13 3 NMAYQQSTL 11 1 FLNMAYQQS 7

TABLE XXIX-V7C HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 179 EQKSKHCMF 28 42 QQDRKIPPL 21 73 GIRNKSSSS 21 165 WMKLETIIL 21 27 ILRGGLSEI 20 181 KSKHCMFSL 20 5 ILDLSVEVL 19 15 SPAAAWKCL 19 113 ANSWRNPVL 19 129 PLWEFLLRL 18 148 SLAFTSWSL 18 102 PPESPDRAL 17 109 ALKAANSWR 17 163 GTWMKLETI 17 19 AWKCLGANI 16 31 GLSEIVLPI 16 137 LLKSQAASG 16 24 GANILRGGL 15 171 IILSKLTQE 15 17 AAAWKCLGA 14 141 QAASGTSL 14 134 LLRLLKSQA 13

TABLE XXIX-V8 HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 FLEEGMGGT 9 5 LEEGMGGTI 6

TABLE XXIX-V13 HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 SPKSLSETF 24 9 FLPNGINGI 14 2 PKSLSETFL 11

TABLE XXIX-V14 HLA-B08-9mers-998P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 NLPLRLFTF 21 3 PLRLFTFWR 13

TABLE XXIX-V21 HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 6 EQKTKHCMF 28 8 KTKHCMFSL 20 4 TQEQKTKHC 11

TABLE XXIX-V25 HLA-B08-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 SQKLKRIKK 23 6 CISQKLKRI 21 3 FLPCISQKL 17

TABLE XXIX-V1 HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 93 IHREHYTSL 23 96 EHYTSLWDL 21 105 RHLLVGKIL 20 315 VHVAYSLCL 20 200 EIENLPLRL 15 426 FYTPPNFVL 15 436 LVLPSIVIL 15 54 YHVVIGSRN 14 264 ITLLSLVYL 14 360 YISFGIMSL 14 365 IMSLGLLSL 14 395 QSTLGYVAL 14 77 THHEDALTK 13 99 TSLWDLRHL 13 125 YPESNAEYL 13 173 QARQQVIEL 13 177 QVIELARQL 13 236 IHPYARNQQ 13 261 IVAITLLSL 13 297 TWLQCRKQL 13 390 EFSFIQSTL 13 428 TPPNFVLAL 13 430 PNFVLALVL 13 5 SMMGSPKSL 12 37 GSGDFAKSL 12 41 FAKSLTIRL 12 71 PHVVDVTHH 12 78 HHEDALTKT 12 100 SLWDLRHLL 12 128 SNAEYLASL 12 133 LASLFPDSL 12 146 FNVVSAWAL 12 196 SSAREIENL 12 214 GPVVVAISL 12 220 ISLATFFFL 12 251 PIEIVNKTL 12 258 TLPIVAITL 12 259 LPIVAITLL 12 287 KYRRFPPWL 12 324 PMRRSERYL 12 326 RRSERYLFL 12 396 STLGYVALL 12 403 LLISTFHVL 12 438 LPSIVILDL 12 441 IVILDLLQL 12 10 PKSLSETCL 11 75 DVTHHEDAL 11 148 VVSAWALQL 11 184 QLNFIPIDL 11 198 AREIENLPL 11 201 IENLPLRLF 11 203 NLPLRLFTL 11 267 LSLVYLAGL 11 274 GLLAAAYQL 11 283 YYGTKYRRF 11 300 QCRKQLGLL 11 341 VHANIENSW 11 351 EEEVWRIEM 11 366 MSLGLLSLL 11 378 SIPSVSNAL 11 383 SNALNWREF 11 411 LIYGWKRAF 11 439 PSIVILDLL 11

TABLE XXIX-V2 HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 SGSPGLQAL 15 35 PPCPADFFL 12 5 GLQALSLSL 11 15 SGFTPFSCL 11 3 SPGLQALSL 10 17 FTPFSCLSL 10 33 CPPPCPADF 9 12 SLSSGFTPF 8 37 CPADFFLYF 8 34 PPPCPADFF 7

TABLE XXIX-V5A HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 NLPLRLFTF 7 8 TFWRGPVVV 7 9 FWRGPVVVA 7 7 FTFWRGPVV 3

TABLE XXIX-V5B HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 19 ELELEFVFL 14 12 SFADTQTEL 13 14 ADTQTELEL 12 20 LELEFVFLL 12 22 LEFVFLLTL 12 23 EFVFLLTLL 11 18 TELELEFVF 10 24 FVFLLTLLL 10 16 TQTELELEF 9 2 REFSFIQIF 7 5 SFIQIFCSF 7

TABLE XXIX-V6 HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 44 PHVSPERVT 15 5 IVILGKIIL 14 7 ILGKIILFL 14 14 FLPCISRKL 12 27 KGWEKSQFL 11 46 VSPERVTVM 10 6 VILGKIILF 8 26 KKGWEKSQF 7 45 HVSPERVTV 7

TABLE XXIX-V7A HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 PKSLSETFL 11 1 SPKSLSETF 7

TABLE XXIX-V7B HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 QSTLGYVAL 14 3 NMAYQQSTL 12 9 STLGYVALL 12

TABLE XXIX-V7C HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 122 PHTNGVGPL 22 5 ILDLSVEVL 15 102 PPESPDRAL 15 113 ANSWRNPVL 14 126 GVGPLWEFL 13 129 PLWEFLLRL 13 130 LWEFLLRLL 13 24 GANILRGGL 12 29 RGGLSEIVL 12 42 QQDRKIPPL 12 50 LSTPPPPAM 12 141 QAASGTLSL 12 160 LGSGTWMKL 12 15 SPAAAWKCL 11 20 WKCLGANIL 11 139 KSQAASGTL 11 148 SLAFTSWSL 11 152 TSWSLGEFL 11 181 KSKHCMFSL 11 127 VGPLWEFLL 10 165 WMKLETIIL 10 168 LETIILSKL 10 183 KHCMFSLIS 10

TABLE XXIX-V8 HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 KSQFLEEGM 6 4 FLEEGMGGT 4 8 GMGGTIPHV 4 5 LEEGMGGTI 3 7 EGMGGTIPH 3 9 MGGTIPHVS 3 6 EEGMGGTIP 2

TABLE XXIX-V13 HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 PKSLSETFL 11 1 SPKSLSETF 7

TABLE XXIX-V14 HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 NLPLRLFTF 7 8 TFWRGPVVV 7 9 FWRGPVVVA 7 7 FTFWRGPVV 3

TABLE XXIX-V21 HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 KTKHCMFSL 11 5 QEQKTKHCM 8 6 EQKTKHCMF 7 4 TQEQKTKHC

TABLE XXIX-V25 HLA-B1510-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 3 FLPCISQKL 10 7 ISQKLKRIK 6 6 CISQKLKRI 4

TABLE XXX-V1 HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 326 RRSERYLFL 26 424 YRFYTPPNF 26 355 WRIEMYISF 25 198 AREIENLPL 24 240 ARNQQSDFY 22 325 MRRSERYLF 22 47 IRLIRGGYH 21 50 IRCGYHVVI 21 104 LRHLLVGKI 21 289 RRFPPWLET 21 416 KRAFEEEYY 21 212 WRGPVVVAI 20 302 RKQLGLLSF 20 417 RAFEEEYYR 20 28 ARKVTVGVI 19 61 RNPKFASEF 19 182 ARQLNFIPI 19 199 REIENLPLR 19 249 KIPIEIVNK 19 303 KQLGLLSFF 19 53 GYHVVIGSR 18 105 RHLLVGKIL 18 179 IELARQLNF 18 214 GPVVVAISL 18 241 RNQQSDFYK 18 274 GLLAAAYQL 18 282 LYYGTKYRR 18 436 LVLPSIVIL 18 21 GINGIKDAR 17 174 ARQQVIELA 17 223 ATFFFLYSF 17 259 LPIVAITLL 17 264 ITLLSLVYL 17 330 RYLFLNMAY 17 360 YISFGIMSL 17 365 IMSLGLLSL 17 366 MSLGLLSLL 17 400 YVALLISTF 17 430 PNFVLALVL 17 441 IVILDLLQL 17 22 INGIKDARK 16 39 GDFAKSLTI 16 40 DFAKSLTIR 16 43 KSLTIRLIR 16 56 VVIGSRNPK 16 112 ILIDVSNNM 16 175 RQQVIELAR 16 177 QVIELARQL 16 196 SSAREIENL 16 206 LRLFTLWRG 16 218 VAISLATFF 16 225 FFFLYSFVR 16 233 RDVIHPYAR 16 313 AMVHVAYSL 16 319 YSLCLPMRR 16 396 STLGYVALL 16 418 AFEEEYYRF 16 443 ILDLLQLCR 16 37 GSGDFAKSL 15 82 ALTKTNIIF 15 87 NIIFVAIHR 15 93 IHREHYTSL 15 96 EHYTSLWDL 15 155 QLGPKDASR 15 173 QARQQVIEL 15 295 LETWLQCRK 15 297 TWLQCRKQL 15 329 ERYLFLNMA 15 390 EFSFIQSTL 15 401 VALLISTFH 15 409 HVLIYGWKR 15 411 LIYGWKRAF 15 438 LPSIVILDL 15 5 SMMGSPKSL 14 10 PKSLSETCL 14 18 LPNGINGIK 14 33 VGVIGSGDF 14 41 FAKSLTIRL 14 57 VIGSRNPKF 14 60 SRNPKFASE 14 77 THHEDALTK 14 120 MRINQYPES 14 128 SNAEYLASL 14 136 LFPDSLIVK 14 146 FNVVSAWAL 14 162 SRQVYICSN 14 167 ICSNNIQAR 14 193 GSLSSAREI 14 200 EIENLPLRL 14 201 IENLPLRLF 14 217 VVAISLATF 14 258 TLPIVAITL 14 261 IVAITLLSL 14 263 AITLLSLVY 14 267 LSLVYLAGL 14 280 YQLYYGTKY 14 281 QLYYGTKYR 14 299 LQCRKQLGL 14 301 CRKQLGLLS 14 308 LSFFFAMVH 14 318 AYSLCLPMR 14 363 FGIMSLGLL 14 392 SFIQSTLGY 14 395 QSTLGYVAL 14 426 FYTPPNFVL 14 439 PSIVILDLL 14 444 LDLLQLCRY 14 35 VIGSGDFAK 13 98 YTSLWDLRH 13 99 TSLWDLRHL 13 103 DLRHLLVGK 13 113 LIDVSNNMR 13 117 SNNMRINQY 13 124 QYPESNAEY 13 129 NAEYLASLF 13 138 PDSLIVKGF 13 148 VVSAWALQL 13 151 AWALQLGPK 13 191 DLGSLSSAR 13 203 NLPLRLFTL 13 220 ISLATFFFL 13 229 YSFVRDVIH 13 239 YARNQQSDF 13 246 DFYKIPIEI 13 251 PIEIVNKTL 13 268 SLVYLAGLL 13 279 AYQLYYGTK 13 283 YYGTKYRRF 13 287 KYRRFPPWL 13 291 FPPWLETWL 13 300 QCRKQLGLL 13 304 QLGLLSFFF 13 306 GLLSFFFAM 13 315 VHVAYSLCL 13 337 AYQQVHANI 13 348 SWNEEEVWR 13 371 LSLLAVTSI 13 378 SIPSVSNAL 13 388 WREFSFIQS 13 403 LLISTFHVL 13 408 FHVLIYGWK 13 435 ALVLPSIVI 13 17 CLPNGINGI 12 70 FPHVVDVTH 12 71 PHVVDVTHH 12 80 EDALTKTNI 12 86 TNIIFVAIH 12 89 IFVAIHREH 12 106 HLLVGKILI 12 114 IDVSNNMRI 12 133 LASLFPDSL 12 134 ASLPFDSLI 12 164 QVYICSNNI 12 184 QLNFIPIDL 12 187 FIPIDLGSL 12 205 PLRLFTLWR 12 219 AISLATFFF 12 231 FVRDVIHPY 12 232 VRDVIHPYA 12 256 NKTLPIVAI 12 272 LAGLLAAAY 12 288 YRRFPPWLE 12 317 VALSLCLPM 12 322 CLPMRRSER 12 328 SERYLFLNM 12 349 WNEEEVWRI 12 352 EEVWRIEMY 12 362 SFGIMSLGL 12 381 SVSNALNWR 12 383 SNALNWREF 12 385 ALNWREFSF 12 428 TPPNFVLAL 12

TABLE XXX-V2 HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 GLQALSLSL 17 9 LSLSLSSGF 15 15 SGFTPFSCL 15 1 SGSPGLQAL 14 3 SPGLQALSL 14 12 SLSSGFTPF 14 23 LSLPSSWDY 14 17 FTPFSCLSL 13 31 YRCPPPCPA 12 33 CPPPCPADF 12 34 PPPCPADFF 12 35 PPCPADFFL 12 24 SLPSSWDYR 11 37 CPADFFLYF 11 2 GSPGLQALS 9 36 PCPADFFLY 8

TABLE XXX-V5A HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 LRLFTFWRG 15 1 NLPLRLFTF 13 3 PLRLFTFWR 11 5 RLFTFWRGP 7

TABLE XXX-V5B HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 REFSFIQIF 20 1 WREFSFIQI 19 5 SFIQIFCSF 16 22 LEFVFLLTL 16 24 FVFLLTLLL 16 12 SFADTQTEL 15 14 ADTQTELEL 15 18 TELELEFVF 15 23 EFVFLLTLL 15 16 TQTELELEF 14 20 LELEFVFLL 14 19 ELELEFVFL 13

TABLE XXX-V6 HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 23 KRIKKGWEK 29 19 SRKLKRIKK 25 6 VILGKIILF 19 13 LFLPCISRK 19 5 IVILGKIIL 18 7 ILGKIIIFL 18 12 ILFLPCISR 18 16 PCISRKLKR 16 26 KKGWEKSQF 16 2 LPSIVILGK 15 18 ISRKLKRIK 15 27 KGWEKSQFL 15 37 EGIGGTIPH 15 14 FLPCISRKL 14 42 TIPHVSPER 14

TABLE XXX-V7A HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 PKSLSETFL 14 1 SPKSLSETF 13 9 FLPNGINGI 12 6 SETFLPNGI 8 7 ETFLPNGIN 6 8 TFLPNGING 6

TABLE XXX-V7B HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 STLGYVALL 16 3 NMAYQQSTL 14 8 QSTLGYVAL 14 5 AYQQSTLGY 13 4 MAYQQSTLG 7

TABLE XXX-V7C HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 21 KCLGANILR 18 29 RGGLSEIVL 18 69 AQESGIRNK 18 167 KLETIILSK 18 175 KLTQEQKSK 18 74 IRNKSSSSS 17 125 NGVGPLWEF 17 128 GPLWEFLLR 17 107 DRALKAANS 16 131 WEFLLRLLK 16 5 ILDLSVEVL 15 20 WKCLGANIL 15 37 LPIEWQQDR 15 42 QQDRKIPPL 15 67 AEAQESGIR 15 100 IDPPESPDR 15 126 GVGPLWEFL 15 129 PLWEFLLRL 15 135 LRLLKSQAA 15 158 EFLGSGTWM 15 160 LGSGTWMKL 15 168 LETIILSKL 15 24 GANILRGGL 14 27 ILRGGLSEI 14 28 LRGGLSEIV 14 38 PIEWQQDRK 14 113 ANSWRNPVL 14 116 WRNPVLPHT 14 139 KSQAASGTL 14 141 QAASGTLSL 14 143 ASGTLSLAF 14 173 LSKLTQEQK 14 13 LASPAAAWK 13 31 GLSEIVLPI 13 44 DRKIPPLST 13 109 ALKAANSWR 13 122 PHTNGVGPL 13 148 SLAFTSWSL 13 151 FTSWSLGEF 13 159 FLGSGTWMK 13 165 WMKLETIIL 13 176 LTQEQKSKH 13 181 KSKHCMFSL 13 39 IEWQQDRKI 12 102 PPESPDRAL 12 103 PESPDRALK 12 130 LWEFLLRLL 12 136 RLLKSQAAS 12 163 GTWMKLETI 12 178 QEQKSKHCM 12 19 AWKCLGANI 11 45 RKIPPLSTP 11 50 LSTPPPPAM 11 108 RALKAANSW 11 115 SWRNPVLPH 11 127 VGPLWEFLL 11 152 TSWSLGEFL 11 157 GEFLGSGTW 11 164 TWMKLETII 11 179 EQKSKHCMF 11 15 SPAAAWKCL 10 30 GGLSEIVLP 10 76 NKSSSSSQI 10 92 EDDEAQDSI 10 75 RNKSSSSSQ 8 85 PVVGVVTED 8 145 GTLSLAFTS 8 171 IILSKLTQE 8 185 CMFSLISGS 8

TABLE XXX-V8 HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 7 EGMGGTIPH 13 1 KSQFLEEGM 11 5 LEEGMGGTI 9 8 GMGGTIPHV 9

TABLE XXX-V13 HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 PKSLSETFL 14 1 SPKSLSETF 13 9 FLPNGINGI 12 6 SETFLPNGI 8 7 ETFLPNGIN 6 8 TFLPNGING 6

TABLE XXX-V14 HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 LRLFTFWRG 15 1 NLPLRLFTF 13 3 PLRLFTFWR 11 5 RLFTFWRGP 7

TABLE XXX-V21 HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 KLTQEQKTK 18 3 LTQEQKTKH 14 8 KTKHCMFSL 13 5 QEQKTKHCM 11 6 EQKTKHCMF 11

TABLE XXX-V25 HLA-B2705-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 LFLPCISQK 18 5 PCISQKLKR 16 7 ISQKLKRIK 15 8 SQKLKRIKK 15 3 FLPCISQKL 14 4 LPCISQKLK 13 6 CISQKLKRI 12 9 QKLKRIKKG 9 1 ILFLPCISQ 8

TABLE XXXI-V1 HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 326 RRSERYLFL 25 198 AREIENLPL 22 424 YRFYTPPNF 22 212 WRGPVVVAI 21 28 ARKVTVGVI 20 50 IRCGYHVVI 20 325 MRRSERYLF 20 104 LRHLLVGKI 19 182 ARQLNFIPI 19 355 WRIEMYISF 19 274 GLLAAAYQL 18 289 RRFPPWLET 18 105 RHLLVGKIL 16 193 GSLSSAREI 15 214 GPVVVAISL 15 441 IVILDLLQL 15 37 GSGDFAKSL 14 39 GDFAKSLTI 14 48 RLIRCGYHV 14 264 ITLLSLVYL 14 306 GLLSFFFAM 14 313 AMVHVAYSL 14 425 RFYTPPNFV 14 430 PNFVLALVL 14 436 LVLPSIVIL 14 47 IRLIRCGYH 13 61 RNPKFASEF 13 68 EFFPHVVDV 13 99 TSLWDLRHL 13 135 SLFPDSLIV 13 148 VVSAWALQL 13 177 QVIELARQL 13 179 IELARQLNF 13 206 LRLFTLWRG 13 220 ISLATFFFL 13 287 KYRRFPPWL 13 297 TWLQCRKQL 13 302 RKQLGLLSF 13 396 STLGYVALL 13 41 FAKSLTIRL 12 85 KTNIIFVAI 12 96 EHYTSLWDL 12 114 IDVSNNMRI 12 120 MRINQYPES 12 125 YPESNAEYL 12 146 FNVVSAWAL 12 157 GPKDASRQV 12 159 KDASRQVYI 12 200 EIENLPLRL 12 223 ATFFFLYSF 12 227 FLYSFVRDV 12 232 VRDVIHPYA 12 261 IVAITLLSL 12 267 LSLVYLAGL 12 268 SLVYLAGLL 12 303 KQLGLLSFF 12 315 VHVAYSLCL 12 317 VAYSLCLPM 12 329 ERYLFLNMA 12 365 IMSLGLLSL 12 366 MSLGLLSLL 12 395 QSTLGYVAL 12 403 LLISTFHVL 12 416 KRAFEEEYY 12 426 FYTPPNFVL 12 428 TPPNFVLAL 12 439 PSIVILDLL 12

TABLE XXXI-V2 HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 GLQALSLSL 14 3 SPGLQALSL 12 15 SGFTPFSCL 12 1 SGSPGLQAL 11 9 LSLSLSSGF 11 17 FTPFSCLSL 11 31 YRCPPPCPA 11 35 PPCPADFFL 11 12 SLSSGFTPF 9 33 CPPPCPADF 9 34 PPPCPADFF 9 37 CPADFFLYF 9 32 RCPPPCPAD 6

TABLE XXXI-V5A HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 LRLFTFWRG 13 7 FTFWRGPVV 11 6 LFTFWRGPV 9 8 TFWRGPVVV 9 1 NLPLRLFTF 8 5 RLFTFWRGP 6

TABLE XXXI-V5B HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 WREFSFIQI 19 2 REFSFIQIF 15 14 ADTQTELEL 13 20 LELEFVFLL 13 22 LEFVFLLTL 13 24 FVFLLTLLL 13 19 ELELEFVFL 11 23 EFVFLLTLL 11 5 SFIQIFCSF 10 12 SFADTQTEL 10 16 TQTELELEF 10 18 TELELEFVF 10

TABLE XXXI-V6 HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 7 ILGKILLFL 13 23 KRIKKGWEK 13 5 IVILGKIIL 12 10 KIILFLPCI 12 27 KGWEKSQFL 12 38 GIGGTIPHV 12 14 FLPCISRKL 11 26 KKGWEKSQF 11 3 PSIVILGKI 10 6 VILGKIILF 10 19 SRKLKRIKK 10 31 KSQFLEEGI 10 43 IPHVSPERV 10 45 HVSPERVTV 10 4 SIVILGKII 9 17 CISRKLKRI 9 35 LEEGIGGTI 9 46 VSPERVTVM 9 20 RKLKRIKKG 6 41 GTIPHVSPE 6

TABLE XXXI-V7A HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 PKSLSETFL 10 1 SPKSLSETF 9 6 SETFLPNGI 9 9 FLPNGINGI 8 3 KSLSETFLP 5 8 TFLPNGING

TABLE XXXI-V7B HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 STLGYVALL 13 8 QSTLGYVAL 12 3 NMAYQQSTL 10 6 YQQSTLGYV 9

TABLE XXXI-V7C HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 28 LRGGLSEIV 18 29 RGGLSEIVL 14 31 GLSEIVLPI 14 126 GVGPLWEFL 14 24 GANILRGGL 13 5 ILDLSVEVL 12 107 DRALKAANS 12 113 ANSWRNPVL 12 116 WRNPVLPHT 12 122 PHTNGVGPL 12 129 PLWEFLLRL 12 135 LRLLKSQAA 12 139 KSQAASGTL 12 141 QAASGTLSL 12 168 LETIILSKL 12 181 KSKHCMFSL 12 4 VILDLSVEV 11 20 WKCLGANIL 11 42 QQDRKIPPL 11 44 DRKIPPLST 11 50 LSTPPPPAM 11 74 IRNKSSSSS 11 82 SQIPVVGVV 11 102 PPESPDRAL 11 119 PVLPHTNGV 11 152 TSWSLGEFL 11 163 GTWMKLETI 11 2 SIVILDLSV 10 15 SPAAAWKCL 10 19 AWKCLGANI 10 76 NKSSSSSQI 10 79 SSSSQIPVV 10 81 SSQIPVVGV 10 112 AANSWRNPV 10 127 VGPLWEFLL 10 130 LWEFLLRLL 10 143 ASGTLSLAF 10 148 SLAFTSWSL 10 158 EFLGSGTWM 10 160 LGSGTWMKL 10 165 WMKLETIIL 10 27 ILRGGLSEI 9 39 IEWQQDRKI 9 78 SSSSSQIPV 9 125 NGVGPLWEF 9 179 EQKSKHCMF 9 66 TAEAQESGI 8 92 EDDEAQDSI 8 151 FTSWSLGEF 8 164 TWMKLETII 8 178 QEQKSKHCM 8 182 SKHCMFSLI 8

TABLE XXXI-V8 HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 GMGGTIPHV 12 1 KSQFLEEGM 10 5 LEEGMGGTI 8

TABLE XXXI-V13 HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 PKSLSETFL 10 1 SPKSLSETF 9 6 SETFLPNGI 9 9 FLPNGINGI 8 3 KSLSETFLP 5 8 TFLPNGING 4

TABLE XXXI-V14 HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 LRLFTFWRG 13 7 FTFWRGPVV 11 6 LFTFWRGPV 9 8 TFWRGPVVV 9 1 NLPLRLFTF 8 5 RLFTFWRGP 6

TABLE XXXI-V21 HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 8 KTKHCMFSL 12 5 QEQKTKHCM 8 6 EQKTKHCMF 8 9 TKHGMFSLI 8

TABLE XXXI-V25 HLA-B2709-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 3 FLPCISQKL 11 6 CISKLKRI 9 2 LFLPCISQK 4

TABLE XXXII-V1 HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 352 EEVWRIEMY 26 201 IENLPLRLF 24 179 IELARQLNF 23 14 SETGLPNGI 21 419 FEEEYYRFY 21 357 IEMYISEGI 20 42 AKSLTIRLI 18 436 LVLPSIVIL 18 117 SNNMRINQY 17 144 KGFNVVSAW 17 259 LPIVAITLL 17 441 IVILDLLQL 17 5 SMMGSPKSL 16 138 PDSLIVKGF 16 177 QVIELARQL 16 199 REIENLPLR 16 203 NLPLRLFTL 16 219 AISLATFFF 16 223 ATFFFLYSF 16 256 NKTLPIVAI 16 263 AITLLSLVY 16 290 RFPPWLETW 16 392 SFIQSTLGY 16 403 LLISTFHVL 16 428 TPPNFVLAL 16 439 PSIVILDLL 16 67 SEFFPHVVD 15 79 HEDALTKTN 15 100 SLWDLRHLL 15 130 AEYLASLFP 15 182 ARQLNFIPI 15 196 SSAREIENL 15 200 EIENLPLRL 15 212 WRGPVVVAI 15 231 FVRDVIHPY 15 252 IEIVNKTLP 15 297 TWLQCRKQL 15 363 FGIMSLGLL 15 378 SIPSVSNAL 15 389 REFSFIQST 15 390 EFSFIQSTL 15 396 STLGYVALL 15 400 YVALLISTF 15 421 EEYYRFYTP 15 430 PNFVLALVL 15 438 LPSIVILDL 15 17 CLPNGINGI 14 37 GSGDFAKSL 14 82 ALTKTNIIF 14 85 KTNIIFVAI 14 96 EHYTSLWDL 14 105 RHLLVGKIL 14 148 VVSAWALQL 14 198 AREIENLPL 14 204 LPLRLFTLW 14 218 VAISLATFF 14 221 SLATFFFLY 14 258 TLPIVAITL 14 264 ITLLSLVYL 14 272 LAGLLAAAY 14 303 KQLGLLSFF 14 313 AMVHVAYSL 14 351 EEEVWRIEM 14 355 WRIEMYISF 14 360 YISFGIMSL 14 365 IMSLGLLSL 14 366 MSLGLLSLL 14 383 SNALNWREF 14 385 ALNWREFSF 14 395 QSTLGYVAL 14 411 LIYGWKRAF 14 426 FYTPPNFVL 14 435 ALVLPSIVI 14 28 ARKVTVGVI 13 46 TIRLIRCGY 13 99 TSLWDLRHL 13 126 PESNAEYLA 13 129 NAEYLASLF 13 133 LASLFPDSL 13 134 ASLFPDSLI 13 146 FNVVSAWAL 13 158 PKDASRQVY 13 180 ELARQLNFI 13 184 QLNFIPIDL 13 240 ARNQQSDFY 13 251 PIEIVNKTL 13 253 EIVNKTLPI 13 268 SLVYLAGLL 13 274 GLLAAAYQL 13 286 TKYRRFPPW 13 287 KYRRFPPWL 13 302 RKQLGLLSF 13 311 FFAMVHVAY 13 323 LPMRRSERY 13 326 RRSERYLFL 13 328 SERYLFLNM 13 330 RYLFLNMAY 13 341 VHANIENSW 13 347 NSWNEEEVW 13 380 PSVSNALNW 13 407 TFHVLIYGW 13 418 AFEEEYYRF 13 420 EEEYYRFYT 13 424 YRFYTPPNF 13 444 LDLLQLCRY 13 10 PKSLSETCL 12 39 GDFAKSLTI 12 41 FAKSLTIRL 12 57 VIGSRNPKF 12 61 RNPKFASEF 12 75 DVTHHEDAL 12 81 DALTKTNII 12 94 HREHYTSLW 12 125 YPESNAEYL 12 128 SNAEYLASL 12 173 QARQQVIEL 12 187 FIPIDLGSL 12 214 GPVVVAISL 12 217 VVAISLATF 12 220 ISLATFFFL 12 261 IVAITLLSL 12 267 LSLVYLAGL 12 280 YQLYYGTKY 12 283 YYGTKYRRF 12 299 LQCRKQLGL 12 300 QCRKQLGLL 12 324 PMRRSERYL 12 325 MRRSERYLF 12 350 NEEEVWRIE 12 353 EVWRIEMYI 12 362 SFGIMSLGL 12 404 LISTFHVLI 12 405 ISTFHVLIY 12

TABLE XXXII-V2 HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 SGSPGLQAL 18 15 SGFTPFSCL 15 33 CPPPCPADF 15 3 SPGLQALSL 14 23 LSLPSSWDY 14 12 SLSSGFTPF 13 21 SCLSLPSSW 13 35 PPCPADFFL 13 36 PCPADFFLY 13 37 CPADFFLYF 13 17 FTPFSCLSL 12 34 PPPCPADFF 12 5 GLQALSLSL 11 9 LSLSLSSGF 11

TABLE XXXII-V5A HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 NLPLRLFTF 16 2 LPLRLFTFW 13

TABLE XXXII-V5B HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 REFSFIQIF 25 22 LEFVFLLTL 25 20 LELEFVFLL 23 18 TELELEFVF 22 5 SFIQIFCSF 16 24 FVFLLTLLL 16 19 ELELEFVFL 15 14 ADTQTELEL 14 23 EFVFLLTLL 14 12 SFADTQTEL 12

TABLE XXXII-V6 HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 35 LEEGIGGTI 21 6 VILGKIILF 17 5 IVILGKIIL 15 7 ILGKIILFL 15 21 KLKRIKKGW 15 3 PSIVILGKI 14 10 KIILFLPCI 14 14 FLPCISRKL 14 17 CISRKLKRI 13 26 KKGWEKSQF 12 29 WEKSQFLEE 12 36 EEGIGGTIP 12 4 SIVILGKII 11 27 KGWEKSQFL 11

TABLE XXXII-V7A HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 6 SETFLPNGI 21 9 FLPNGNGI 14 1 SPKSLSETF 12 2 PKSLSETFL 12

TABLE XXXII-V7B HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 AYQQSTLGY 15 9 STLGYVALL 15 8 QSTLGYVAL 14 3 NMAYQQSTL 12

TABLE XXXII-V7C HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 33 SEIVLPIEW 26 157 GEFLGSGTW 24 168 LETIILSKL 23 39 IEWQQDRKI 20 143 ASGTLSLAF 17 51 STPPPPAMW 16 70 QESGIRNKS 16 103 PESPDRALK 16 113 ANSWRNPVL 16 131 WEFLLRLLK 16 42 QQDRKIPPL 15 5 ILDLSVEVL 14 61 EEAGATAEA 14 10 VEVLASPAA 13 12 VLASPAAAW 13 15 SPAAAWKCL 13 20 WKCLGANIL 13 29 RGGLSEIVL 13 60 TEEAGATAE 13 67 AEAQESGIR 13 91 TEDDEAQDS 13 102 PPESPDRAL 13 108 RALKAANSW 13 125 NGVGPLWEF 13 126 GVGPLWEFL 13 127 VGPLWEFLL 13 130 LWEFLLRLL 13 146 TLSLAFTSW 13 160 LGSGTWMKL 13 165 WMKLETIIL 13 31 GLSEIVLPI 12 122 PHTNGVGPL 12 123 HTNGVGPLW 12 129 PLWEFLLRL 12 139 KSQAASGTL 12 141 QAASGTLSL 12 151 FTSWSLGEF 12 179 EQKSKHCMF 12

TABLE XXXII-V8 HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 LEEGMGGTI 20 6 EEGMGGTIP 12

TABLE XXXII-V13 HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 6 SETFLPNGI 21 9 FLPNGINGI 14 1 SPKSLSETF 12 2 PKSLSETFL 12

TABLE XXXII-V14 HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 1 NLPLRLFTF 16 2 LPLRLFTFW 13

TABLE XXXII-V21 HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 6 EQKTKHCMF 13 5 QEQKTKHCM 11 8 KTKHCMFSL 11 9 TKHCMFSLI 10

TABLE XXXII-V25 HLA-B4402-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 3 FLPCISQKL 13 6 CIDQKLKRI 12 2 LFLPCISQK 8 9 QKLKRIKKG 8

TABLE XXXIIII-V1 HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 81 DALTKTNII 29 27 DARKVTVGV 26 65 FASEFFPHV 23 374 LAVTSIPSV 23 434 LALVLPSIV 23 438 LPSIVILDL 22 246 DFYKIPIEI 21 262 VAITLLSLV 21 368 LGLLSLLAV 21 428 TPPNFVLAL 21 429 PPNFVLALV 21 23 NGIKDARKV 20 157 GPKDASRQV 20 214 GPVVVAISL 20 259 LPIVAITLL 20 41 FAKSLTIRL 19 125 YPESNAEYL 19 133 LASLFPDSL 19 173 QARQQVIEL 19 250 IPIEIVNKT 19 291 FPPWLETWL 19 50 IRGGYHVVI 18 228 LYSFVRDVI 17 336 MAYQQVHAN 17 371 LSLLAVTSI 17 28 ARKVTVGVI 16 39 GDFAKSLTI 16 70 FPHVVDVTH 16 104 LRHLLVGKI 16 141 LIVKGFNVV 16 160 DASRQVYIC 16 204 LPLRLFTLW 16 227 FLYSFVRDV 16 237 HPYARNQQS 16 317 VAYSLCLPM 16 52 CGYHVVIGS 15 137 FPDSLIVKG 15 164 QVYICSNNI 15 171 NIQARQQVI 15 193 GSLSSAREI 15 210 TLWRGPVVV 15 212 WRGPVVVAI 15 276 LAAAYQLYY 15 349 WNEEEVWRI 15 363 FGIMSLGLL 15 397 TLGYVALLI 15 425 RFYTPPNFV 15 18 LPNGINGIK 14 25 IKDARKVTV 14 114 IDVSNNMRI 14 152 WALQLGPKD 14 209 FTLWRGPVV 14 222 LATFFFLYS 14 242 NQQSDFYKI 14 258 TLPIVAITL 14 278 AAYQLYYGT 14 379 IPSVSNALN 14 386 LNWREFSFI 14 398 LGYVALLIS 14 401 VALLISTFH 14 404 LISTFHVLI 14 433 VLALVLPSI 14 435 ALVLPSIVI 14

TABLE XXXIIII-V2 HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 3 SPGLQALSL 18 35 PPCPADFFL 16 15 SGFTPFSCL 15 1 SGSPGLQAL 13 7 QALSLSLSS 13 18 TPFSCLSLP 13 25 LPSSWDYRC 13 37 CPADFFLYF 13 33 CPPPCPADF 12 34 PPPCPADFF 12 17 FTPFSCLSL 10 4 PGLQALSLS 9 5 GLQALSLSL 8

TABLE XXXIIII-V5A HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 LPLRLFTFW 16 8 TFWRGPVVV 15 7 FTFWRGPVV 13 6 LFTFWRGPV 10 9 FWRGPVVVA 8 4 LRLFTFWRG 7

TABLE XXXIIII-V5B HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 20 LELEFVFLL 14 1 WREFSFIQI 13 22 LEFVFLLTL 13 13 FADTQTELE 12 12 SFADTQTEL 9 17 QTELELEFV 9 24 FVFLLTLLL 9 14 ADTQTELEL 8 18 TELELEFVF 8 19 ELELEFVFL 8 23 EFVFLLTLL 8 15 DTQTELELE 6

TABLE XXXIIII-V6 HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 43 IPHVSPERV 23 2 LPSIVILGK 16 27 KGWEKSQFL 16 35 LEEGIGGTI 15 15 LPCISRKLK 14 17 CISRKLKRI 14 3 PSIVILGKI 13 39 IGGTIPHVS 13 38 GIGGTIPHV 12 4 SIVILGKII 11 7 ILGKIILFL 11 10 KIILFLPCI 11 14 FLPCISRKL 11 45 HVSPERVTV 11

TABLE XXXIIII-V7A HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 FLPNGINGI 14 1 SPKSLSETF 12 6 SETFLPNGI 12 2 PKSLSETFL

TABLE XXXIIII-V7B HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 MAYQQSTLG 16 6 YQQSTLGYV 12 9 STLGYVALL 12 3 NMAYQQSTL 9 8 QSTLGYVAL 7

TABLE XXXIIII-V7C HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 66 TAEAQESGI 22 101 DPPESPDRA 20 112 AANSWRNPV 19 15 SPAAAWKCL 18 160 LGSGTWMKL 18 29 RGGLSEIVL 17 84 IPVVGVVTE 17 102 PPESPDRAL 17 141 QAASGTLSL 17 24 GANILRGGL 16 39 IEWQQDRKI 16 31 GLSEIVLPI 15 68 EAQESGIRN 15 82 SQIPVVGVV 15 108 RALKAANSW 15 149 LAFTSWSLG 15 163 GTWMKLETI 15 5 ILDLSVEVL 14 27 ILRGGLSEI 14 37 LPIEWQQDR 14 47 IPPLSTPPP 14 48 PPLSTPPPP 14 54 PPPAMWTEE 14 121 LPHTNGVGP 14 127 VGPLWEFLL 14 128 GPLWEFLLR 14 4 VILDLSVEV 13 13 LASPAAAWK 13 18 AAWKCLGAN 13 52 TPPPPAMWT 13 53 PPPPAMWTE 13 62 EAGATAEAQ 13 95 EAQDSIDPP 13 142 AASGTLSLA 13 164 TWMKLETII 13 17 AAAWKCLGA 12 64 GATAEAQES 12 76 NKSSSSSQI 12 79 SSSSQIPVV 12 92 EDDEAQDSI 12 105 SPDRALKAA 12 111 KAANSWRNP 12 118 NPVLPHTNG 12 129 PLWEFLLRL 12 182 SKHCMFSLI 12 16 PAAAWKCLG 11 28 LRGGLSEIV 11 56 PAMWTEEAG 11 81 SSQIPVVGV 11 119 PVLPHTNGV 11 168 LETIIISKL 11 19 AWKCLGANI 10 23 LGANILRGG 10 30 GGLSEIVLP 10 55 PPAMWTEEA 10 78 SSSSSQIPV 10 113 ANSWRNPVL 10 130 LWEFLLRLL 10

TABLE XXXIIII-V8 HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 5 LEEGMGGTI 16 8 GMGGTIPHV 12 9 MGGTIPHVS 12 7 EGMGGTIPH 8

TABLE XXXIIII-V13 HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 FLPNGINGI 14 1 SPKSLSETF 12 6 SETFLPNGI 12 2 PKSLSETFL 8

TABLE XXXIIII-V14 HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 2 LPLRLFTFW 16 8 TFWRGPVVV 15 7 FTFWRGPVV 13 6 LFTFWRGPV 10 9 FWRGPVVVA 8 4 LRLFTFWRG 7

TABLE XXXIIII-V21 HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 9 TKHCMFSLI 13 3 LTQEQKTKH 7 8 KTKHCMFSL 6

TABLE XXXIIII-V25 HLA-B5101-9mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. Pos 123456789 score 4 LPCISQKLK 14 6 CISQKLKRI 14 3 FLPCISQKL 10 9 QKLKRIKKG 7

TABLE XXXIV-V1 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 351 EEEVWRIEMY 26 391 FSFIQSTLGY 26 418 AFEEEYYRFY 26 443 ILDLLQLCRY 26 220 ISLATFFFLY 24 262 VAITLLSLVY 23 327 RSERYLFLNM 23 45 LTIRLIRCGY 22 275 LLAAAYQLYY 22 404 LISTFHVLIY 22 116 VSNNMRINQY 20 123 NQYPESNAEY 20 271 YLAGLLAAAY 19 279 AYQLYYGTKY 19 427 YTPPNFVLAL 19 38 SGDFAKSLTI 18 274 GLLAAAYQLY 18 101 LWDLRHLLVG 17 157 GPKDASRQVY 17 178 VIELARQLNF 17 230 SFVRDVIHPY 17 239 YARNQQSDFY 17 396 STLGYVALLI 17 66 ASEFFPHVVD 16 89 IFVAIHREHY 16 94 HREHYTSLWD 16 129 NAEYLASLFP 16 310 FFFAMVHVAY 16 322 CLPMRRSERY 16 329 ERYLFLNMAY 16 350 NEEEVWRIEM 15 414 GWKRAFEEEY 15 415 WKRAFEEEYY 15 13 LSETCLPNGI 14 125 YPESNAEYLA 14 244 QSDFYKIPIE 14 257 KTLPIVAITL 14 76 VTHHEDALTK 13 198 AREIENLPLR 13 366 MSLGLLSLLA 13 420 EEEYYRFYTP 13 25 IKDARKVTVG 12 135 SLFPDSLIVK 12 137 FPDSLIVKGF 12 200 EIENLPLRLF 12 221 SLATFFFLYS 12 251 PIEIVNKTLP 12 268 SLVYLAGLLA 12 419 FEEEYYRFYT 12 439 PSIVILDLLQ 12

TABLE XXXIV-V2 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 35 PPCPADFFLY 24 22 CLSLPSSWDY 16 28 SWDYRCPPPC 12 2 GSPGLQALSL 11

TABLE XXXIV-V5A HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 FTFWRGPVVV 8 1 ENLPLRLFTF 4 2 NLPLRLFTFW 4 4 PLRLFTFWRG 4 10 FWRGPVVVAI 3

TABLE XXXIV-V5B HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 14 FADTQTELEL 17 18 QTELELEFVF 17 22 ELEFVFLLTL 17 20 ELELEFVFLL 14 16 DTQTELELEF 12 21 LELEFVFLLT 11 2 WREFSFIQIF 10 5 FSFIQIFCSF 8 24 EFVFLLTLLL 8

TABLE XXXIV-V6 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 29 GWEKSQFLEE 19 35 FLEEGIGGTI 13 36 LEEGIGGTIP 12 1 LVLPSIVILG 11 19 ISRKLKRIKK 11 42 GTIPHVSPER 10 9 LGKIIIFLPC 9

TABLE XXXIV-V7A HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 6 LSETFLPNGI 14 4 KSLSETFLPN 13 8 ETFLPNGING 11

TABLE XXXIV-V7B HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 5 MAYQQSTLGY 21 10 STLGYVALLI 17

TABLE XXXIV-V7C HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 131 LWEFLLRLLK 19 33 LSEIVLPIEW 18 91 VTEDDEAQDS 17 60 WTEEAGATAE 16 100 SIDPPESPDR 16 70 AQESGIRNKS 14 94 DDEAQDSIDP 14 6 ILDLSVEVLA 13 103 PPESPDRALK 13 124 HTNGVGPLWE 13 168 KLETIILSKL 13 10 SVEVLASPAA 12 39 PIEWQQDRKI 12 43 QQDRKIPPLS 12 52 STPPPPAMWT 12 104 PESPDRALKA 12 106 SPDRALKAAN 12 128 VGPLWEFLLR 12 170 ETIILSKLTQ 12 97 AQDSIDPPES 11 115 NSWRNPVLPH 11 154 SWSLGEFLGS 11 2 PSIVILDLSV 10 61 TEEAGATAEA 10 67 TAEAQESGIR 10 92 TEDDEAQDSI 10 93 EDDEAQDSID 10 157 LGEFLGSGTW 10 162 GSGTWMKLET 10 178 TQEQKSKHCM 10 51 LSTPPPPAMW 9 146 GTLSLAFTSW 9 182 KSKHCMFSLI 9

TABLE XXXIV-V8 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 5 FLEEGMGGTI 13 6 LEEGMGGTIP 12

TABLE XXXIV-V13 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 6 LSETFLPNGI 14 4 KSLSETFLPN 13 8 ETFLPNGING 11

TABLE XXXIV-V14 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 FTFWRGPVVV 8 1 ENLPLRLFTF 4 2 NLPLRLFTFW 4 4 PLRLFTFWRG 4 10 FWRGPVVVAI 3

TABLE XXXIV-V21 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 9 KTKHCMFSLI 11 5 TQEQKTKHCM 10 1 LSKLTQEQKT 6 4 LTQEQKTKHC 6 10 TKHCMFSLIS 6

TABLE XXXIV-V25 HLA-A1-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 ISQKLKRIKK 11 5 LPCISQKLKR 8 3 LFLPCISQKL 6

TABLE XXXV-V1 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 373 LLAVTSIPSV 31 266 LLSLVYLAGL 29 107 LLVGKILIDV 28 367 SLGLLSLLAV 28 435 ALVLPSIVIL 28 364 GIMSLGLLSL 27 132 YLASLFPDSL 26 370 LLSLLAVTSI 26 437 VLPSIVILDL 26 82 ALTKTNIIFV 25 100 SLWDLRHLLV 25 140 SLIVKGFNVV 25 263 AITLLSLVYL 25 306 GLLSFFFAMV 25 402 ALLISTFHVL 25 440 SIVILDLLQL 25 258 TLPIVAITLL 24 365 IMSLGLLSLL 24 403 LLISTFHVLI 24 427 YTPPNFVLAL 24 24 GIKDARKVTV 23 48 RLIRCGYHVV 23 103 DLRHLLVGKI 23 433 VLALVLPSIV 23 92 AIHREHYTSL 22 260 PIVAITLLSL 22 261 IVAITLLSLV 22 298 WLQCRKQLGL 22 432 FVLALVLPSI 22 207 RLFTLWRGPV 21 210 TLWRGPVVVA 21 257 KTLPIVAITL 21 385 ALNWREFSFI 21 49 LIRCGYHVVI 20 98 YTSLWDLRHL 20 172 IQARQQVIEL 20 186 NFIPIDLGSL 20 219 AISLATFFFL 20 227 FLYSFVRDVI 20 249 KIPIEIVNKT 20 253 EIVNKTLPIV 20 12 SLSETCLPNG 19 135 SLFPDSLIVK 19 142 IVKGFNVVSA 19 197 SAREIENLPL 19 209 FTLWRGPVVV 19 211 LWRGPVVVAI 19 271 YLAGLLAAAY 19 312 FAMVHVAYSL 19 396 STLGYVALLI 19 16 TCLPNGINGI 18 65 FASEFFPHVV 18 67 SEFFPHVVDV 18 113 LIDVSNNMRI 18 359 MYISFGIMSL 18 392 SFIQSTLGYV 18 106 HLLVGKILID 17 179 IELARQLNFI 17 202 ENLPLRLFTL 17 250 IPIEIVNKTL 17 264 ITLLSLVYLA 17 269 LVYLAGLLAA 17 348 SWNEEEVWRI 17 361 ISFGIMSLGL 17 369 GLLSLLAVTS 17 401 VALLISTFHV 17 26 KDARKVTVGV 16 41 FAKSLTIRLI 16 111 KILIDVSNNM 16 112 ILIDVSNNMR 16 127 ESNAEYLASL 16 195 LSSAREIENL 16 223 ATFFFLYSFV 16 226 FFLYSFVRDV 16 268 SLVYLAGLLA 16 299 LQCRKQLGLL 16 356 RIEMYISFGI 16 362 SFGIMSLGLL 16 377 TSIPSVSNAL 16 428 TPPNFVLALV 16 434 LALVLPSIVI 16 438 LPSIVILDLL 16 443 ILDLLQLCRY 16 27 DARKVTVGVI 15 36 IGSGDFAKSL 15 44 SLTIRLIRCG 15 47 IRLIRCGYHV 15 147 NVVSAWALQL 15 166 YICSNNIQAR 15 189 PIDLGSLSSA 15 199 REIENLPLRL 15 221 SLATFFFLYS 15 255 VNKTLPIVAI 15 273 AGLLAAAYQL 15 275 LLAAAYQLYY 15 314 MVHVAYSLCL 15 335 NMAYQQVHAN 15 336 MAYQQVHANI 15 345 IENSWNEEEV 15 394 IQSTLGYVAL 15 395 QSTLGYVALL 15 404 LISTFHVLIY 15 411 LIYGWKRAFE 15

TABLE XXXV-V2 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 2 GSPGLQALSL 16 5 GLQALSLSLS 15 16 GFTPFSCLSL 15 10 SLSLSSGFTP 14 8 ALSLSLSSGF 13 12 SLSSGFTPFS 13 24 SLPSSWDYRC 13 4 PGLQALSLSL 12 7 QALSLSLSSG 12 14 SSGFTPFSCL 11 22 CLSLPSSWDY 10 9 LSLSLSSGFT 8 17 FTPFSCLSLP 8 6 LQALSLSLSS 7 34 PPPCPADFFL 7

TABLE XXXV-V5A HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 6 RLFTFWRGPV 21 8 FTFWRGPVVV 18 10 FWRGPVVVAI 18 7 LFTFWRGPVV 11 9 TFWRGPVVVA 11 2 NLPLRLFTFW 10

TABLE XXXV-V5B HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 22 ELEFVFLLTL 22 20 ELELEFVFLL 20 14 FADTQTELEL 18 23 LEFVFLLTLL 17 19 TELELEFVFL 16 17 TQTELELEFV 15 12 CSFADTQTEL 13 9 QIFCSFADTQ 11 21 LELEFVFLLT 11 1 NWREFSFIQI 10 7 FIQIFCSFAD 10

TABLE XXXV-V6 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 7 VILGKIILFL 28 35 FLEEGIGGTI 22 5 SIVILGKIIL 20 14 LFLPCISRKL 18 43 TIPHVSPERV 18 2 VLPSIVILGK 17 13 ILFLPCISRK 17 3 LPSIVILGKI 16 8 ILGKIILFLP 16 10 GKIILFLPCI 16 38 EGIGGTIPHV 16 1 LVLPSIVILG 14 46 HVSPERVTVM 14 12 IILFLPCISR 13 34 QFLEEGIGGT 13

TABLE XXXV-V7A HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 5 SLSETFLPNG 19 9 TFLPNGINGI 18 2 SPKSLSETFL 11 6 LSETFLPNGI 11 10 FLPNGINGIK 11

TABLE XXXV-V7B HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 10 STLGYVALLI 19 2 FLNMAYQQST 18 6 AYQQSTLGYV 16 3 LNMAYQQSTL 15 9 QSTLGYVALL 15 8 QQSTLGYVAL 13 4 NMAYQQSTLG 9

TABLE XXXV-V7C HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 5 VILDLSVEVL 26 168 KLETIILSKL 26 27 NILRGGLSEI 24 28 ILRGGLSEIV 24 130 PLWEFLLRLL 24 160 FLGSGTWMKL 23 4 IVILDLSVEV 22 66 ATAEAQESGI 19 81 SSSQIPVVGV 19 156 SLGEFLGSGT 19 6 ILDLSVEVLA 18 32 GLSEIVLPIE 18 112 KAANSWRNPV 18 113 AANSWRNPVL 18 129 GPLWEFLLRL 18 8 DLSVEVLASP 17 19 AAWKCLGANI 17 79 SSSSSQIPVV 17 127 GVGPLWEFLL 17 134 FLLRLLKSQA 17 135 LLRLLKSQAA 17 141 SQAASGTLSL 17 31 GGLSEIVLPI 16 42 WQQDRKIPPL 16 58 AMWTEEAGAT 16 82 SSQIPVVGVV 16 84 QIPVVGVVTE 16 122 LPHTNGVGPL 16 137 RLLKSQAASG 16 138 LLKSQAASGT 16 148 LSLAFTSWSL 16 13 VLASPAAAWK 15 23 CLGANILRGG 15 24 LGANILRGGL 15 152 FTSWSLGEFL 15 163 SGTWMKLETI 15 3 SIVILDLSVE 14 29 LRGGLSEIVL 14 39 PIEWQQDRKI 14 121 VLPHTNGVGP 14 139 LKSQAASGTL 14 142 QAASGTLSLA 14 164 GTWMKLETII 14 171 TIILSKLTQE 14 172 IILSKLTQEQ 14 18 AAAWKCLGAN 13 50 PLSTPPPPAM 13 100 SIDPPESPDR 13 149 SLAFTSWSLG 13 2 PSIVILDLSV 12 20 AWKGLGANIL 12 47 KIPPLSTPPP 12 52 STPPPPAMWT 12 83 SQIPVVGVVT 12 102 DPPESPDRAL 12 119 NPVLPHTNGV 12 126 NGVGPLWEFL 12 144 ASGTLSLAFT 12 173 ILSKLTQEQK 12 176 KLTQEQKSKH 12 181 QKSKHCMFSL 12

TABLE XXXV-V8 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 5 FLEEGMGGTI 22 8 EGMGGTIPHV 15 9 GMGGTIPHVS 12

TABLE XXX-V13 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 5 SLSETFLPNG 19 9 TFLPNGINGI 18 2 SPKSLSETFL 11 6 LSETFLPNGI 11 10 FLPNGINGIK 11

TABLE XXXV-V14 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 6 RLFTFWRGPV 21 8 FTFWRGPVVV 18 10 FWRGPVVVAI 18 7 LFTFWRGPVV 11 9 TFWRGPVVVA 11 2 NLPLRLFTFW 10

TABLE XXXV-V21 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 3 KLTQEQKTKH 12 9 KTKHCMFSLI 12 8 QKTKHCMFSL 11 1 LSKLTQEQKT 7 4 LTQEQKTKHC 7 2 SKLTQEQKTK 5

TABLE XXXV-V25 HLA-A0201-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 3 LFLPCISQKL 18 2 ILFLPCISQK 17 1 IILFLPCISQ 13 4 FLPCISQKLK 10 6 PCISQKLKRI 10 7 CISQKLKRIK 8

TABLE XXXVI-V1 HLA-A0203-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 270 VYLAGLLAAA 27 269 LVYLAGLLAA 19 144 KGFNVVSAWA 18 271 YLAGLLAAAY 17

TABLE XXXVI-V2 HLA-A0203-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 30 DYRCPPPCPA 10 31 YRCPPPCPAD 9 1 SGSPGLQALS 8 32 RCPPPCPADF 8

TABLE XXXVI-V5A HLA-A0203-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 9 TFWRGPVVVA 10 10 FWRGPVVVAI 9

TABLE XXXVI-V5B HLA-A0203-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 6 SFIQIFCSFA 10 7 FIQIFCSFAD 9 8 IQIFCSFADT 8

TABLE XXXVI-V6 HLA-A0203-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XXXVI-V7A HLA-A0203-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XXXVI-V7B HLA-A0203-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 7 YQQSTLGYVA 10 8 QQSTLGYVAL 9 9 QSTLGYVALL 8

TABLE XXXVI-V7C HLA-A0203-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 11 VEVLASPAAA 27 10 SVEVLASPAA 19 105 ESPDRALKAA 19 135 LLRLLKSQAA 19 57 PAMWTEEAGA 18 59 MWTEEAGATA 18 61 TEEAGATAEA 18 12 EVLASPAAAW 17 106 SPDRALKAAN 17 136 LRLLKSQAAS 17

TABLE XXXVI-V8 HLA-A0203-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XXXVI-V13 HLA-A0203-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XXXVI-V14 HLA-A0203-10mers-98P4B6 Pos 1234567890 score 9 TFWRGPVVVA 10 10 FWRGPVVVAI 9

TABLE XXXVI-V21 HLA-A0203-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XXXVI-V25 HLA-A0203-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XXXVII-V1 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 135 SLFPDSLIVK 28 34 GVIGSGDFAK 26 271 YLAGLLAAAY 26 48 RLIRCGYHVV 24 21 GINGIKDARK 23 216 VVVAISLATF 23 369 GLLSLLAVTS 23 17 CLPNGINGIK 22 55 HVVIGSRNPK 22 275 LLAAAYQLYY 22 278 AAYQLYYGTK 22 307 LLSFFFAMVH 22 112 ILIDVSNNMR 21 142 IVKGFNVVSA 21 155 QLGPKDASRQ 21 210 TLWRGPVVVA 21 76 VTHHEDALTK 20 217 VVAISLATFF 20 248 YKIPIEIVNK 20 274 GLLAAAYQLY 20 281 QLYYGTKYRR 20 294 WLETWLQCRK 20 402 ALLISTFHVL 20 2 ESISMMGSPK 19 49 LIRCGYHVVI 19 56 VVIGSRNPKF 19 102 WDLRHLLVGK 19 147 NVVSAWALQL 19 227 FLYSFVRDVI 19 269 LVYLAGLLAA 19 375 AVTSIPSVSN 19 443 ILDLLQLCRY 19 24 GIKDARKVTV 18 140 SLIVKGFNVV 18 333 FLNMAYQQVH 18 410 VLIYGWKRAF 18 411 LIYGWKRAFE 18 435 ALVLPSIVIL 18 442 VILDLLQLCR 18 46 TIRLIRCGYH 17 92 AIHREHYTSL 17 164 QVYICSNNIQ 17 177 QVIELARQLN 17 254 IVNKTLPIVA 17 261 IVAITLLSLV 17 268 SLVYLAGLLA 17 331 YLFLNMAYQQ 17 400 YVALLISTFH 17 403 LLISTFHVLI 17 404 LISTFHVLIY 17 30 KVTVGVIGSG 16 123 NQYPESNAEY 16 141 LIVKGFNVVS 16 178 VIELARQLNF 16 207 RLFTLWRGPV 16 234 DVIHPYARNQ 16 262 VAITLLSLVY 16 263 AITLLSLVYL 16 265 TLLSLVYLAG 16 306 GLLSFFFAMV 16 322 CLPMRRSERY 16 340 QVHANIENSW 16 367 SLGLLSLLAV 16 385 ALNWREFSFI 16 432 FVLALVLPSI 16 433 VLALVLPSIV 16 440 SIVILDLLQL 16 441 IVILDLLQLC 16 32 TVGVIGSGDF 15 100 SLWDLRHLLV 15 106 HLLVGKILID 15 121 RINQYPESNA 15 153 ALQLGPKDAS 15 187 FIPIDLGSLS 15 221 SLATFFFLYS 15 235 VIHPYARNQQ 15 257 KTLPIVAITL 15 260 PIVAITLLSL 15 320 SLCLPMRRSE 15 372 SLLAVTSIPS 15 393 FIQSTLGYVA 15 436 LVLPSIVILD 15 60 SRNPKFASEF 14 88 IIFVAIHREH 14 103 DLRHLLVGKI 14 108 LVGKILIDVS 14 111 KILIDVSNNM 14 132 YLASLFPDSL 14 150 SAWALQLGPK 14 171 NIQARQQVIE 14 180 ELARQLNFIP 14 189 PIDLGSLSSA 14 190 IDLGSLSSAR 14 205 PLRLFTLWRG 14 215 PVVVAISLAT 14 231 FVRDVIHPYA 14 266 LLSLVYLAGL 14 279 AYQLYYGTKY 14 316 HVAYSLCLPM 14 370 LLSLLAVTSI 14 45 LTIRLIRCGY 13 75 DVTHHEDALT 13 82 ALTKTNIIFV 13 128 SNAEYLASLF 13 154 LQLGPKDASR 13 157 GPKDASRQVY 13 166 YICSNNIQAR 13 191 DLGSLSSARE 13 200 EIENLPLRLF 13 204 LPLRLFTLWR 13 240 ARNQQSDFYK 13 298 WLQCRKQLGL 13 304 QLGLLSFFFA 13 310 FFFAMVHVAY 13 314 MVHVAYSLCL 13 321 LCLPMRRSER 13 329 ERYLFLNMAY 13 353 EVWRIEMYIS 13 364 GIMSLGLLSL 13 373 LLAVTSIPSV 13 397 TLGYVALLIS 13 399 GYVALLISTF 13 409 HVLIYGWKRA 13 437 VLPSIVILDL 13 445 DLLQLCRYPD 13

TABLE XXXVII-V2 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 ALSLSLSSGF 21 10 SLSLSSGFTP 19 22 CLSLPSSWDY 17 5 GLQALSLSLS 15 32 RCPPPCPADF 15 12 SLSSGFTPFS 11 24 SLPSSWDYRC 11 2 GSPGLQALSL 10 33 CPPPCPADFF 10

TABLE XXXVII-V5A HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 6 RLFTFWRGPV 16 4 PLRLFTFWRG 14 1 ENLPLRLFTF 13 2 NLPLRLFTFW 12 9 TFWRGPVVVA 11 3 LPLRLFTFWR 10 10 FWRGPVVVAI 10 8 FTFWRGPVVV 9 7 LFTFWRGPVV 7

TABLE XXXVII-V5B HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 9 QIFCSFADTQ 17 22 ELEFVFLLTL 17 18 QTELELEFVF 11 20 ELELEFVFLL 11 7 FIQIFCSFAD 10 8 IQIFCSFADT 8

TABLE XXXVII-V6 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 13 ILFLPCISRK 26 2 VLPSIVILGK 23 15 FLPCISRKLK 21 18 CISRKIKRIK 21 6 IVILGKIILF 20 22 KLKRIKKGWE 19 35 FLEEGIGGTI 19 12 IILFLPCISR 18 46 HVSPERVTVM 18 23 LKRIKKGWEK 17 11 KIILFLPCIS 16 19 ISRKLKRIKK 16 1 LVLPSIVILG 15 7 VILGKIILFL 15 25 RIKKGWEKSQ 15 26 IKKGWEKSQF 15 39 GIGGTIPHVS 15 8 ILGKIILFLP 12

TABLE XXXVII-V7A HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 10 FLPNGINGIK 22 5 SLSETFLPNG 12

TABLE XXXVII-V7B HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 5 MAYQQSTLGY 13 2 FLNMAYQQST 12 10 STLGYVALLI 11 3 LNMAYQQSTL 9 7 YQQSTLGYVA 7 8 QQSTLGYVAL 7 1 LFLNMAYQQS 6 9 QSTLGYVALL 6

TABLE XXXVII-V7C HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 13 VLASPAAAWK 28 173 ILSKLTQEQK 25 137 RLLKSQAASG 24 12 EVLASPAAAW 21 134 FLLRLLKSQA 21 4 IVILDLSVEV 20 36 IVLPIEWQQD 20 120 PVLPHTNGVG 20 176 KLTQEQKSKH 20 83 SQIPVVGVVT 18 84 QIPVVGVVTE 18 156 SLGEFLGSGT 18 167 MKLETIILSK 18 3 SIVILDLSVE 17 6 ILDLSVEVLA 17 28 ILRGGLSEIV 17 74 GIRNKSSSSS 17 90 VVTEDDEAQD 17 121 VLPHTNGVGP 17 138 LLKSQAASGT 17 27 NILRGGLSEI 16 100 SIDPPESPDR 16 110 ALKAANSWRN 16 168 KLETIILSKL 16 171 TIILSKLTQE 16 5 VILDLSVEVL 15 8 DLSVEVLASP 15 26 ANILRGGLSE 15 37 VLPIEWQQDR 15 135 LLRLLKSQAA 15 147 TLSLAFTSWS 15 149 SLAFTSWSLG 15 159 EFLGSGTWMK 15 175 SKLTQEQKSK 15 38 LPIEWQQDRK 14 47 KIPPLSTPPP 14 103 PPESPDRAKL 14 109 RALKAANSWR 14 131 LWEFLLRLLK 14 127 GVGPLWEFLL 13 143 AASGTLSLAF 13

TABLE XXXVII-V8 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 5 FLEEGMGGTI 19

TABLE XXXVII-V13 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 10 FLPNGINGIK 22 5 SLSETFLPNG 12

TABLE XXXVII-V14 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 6 RLFTFWRGPV 16 4 PLRLFTFWRG 14 1 ENLPLRLFTF 13 2 NLPLRLFTFW 12 9 TFWRGPVVVA 11 3 LPLRLFTFWR 10 10 FWRGPVVVAI 10 8 FTFWRGPVVV 9 7 LFTFWRGPVV 7

TABLE XXXVII-V21 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 3 KLTQEQKTKH 18 2 SKLTQEQKTK 17

TABLE XXXVII-V25 HLA-A3-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 2 ILFLPCISQK 29 4 FLPCISQKLK 20 7 CISQKLKRIK 18 1 IILFLPCISQ 14

TABLE XXXVII-V1 HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 216 VVVAISLATF 27 296 ETWLQCRKQL 27 200 EIENLPLRLF 26 147 NVVSAWALQL 25 351 EEEVWRIEMY 25 202 ENLPLRLFTL 24 56 VVIGSRNPKF 23 127 ESNAEYLASL 23 427 YTPPNFVLAL 23 440 SIVILDLLQL 23 45 LTIRLIRCGY 22 234 DVIHPYARNQ 22 253 EIVNKTLPIV 22 260 PIVAITLLSL 22 329 ERYLFLNMAY 21 15 ETCLPNGING 20 32 TVGVIGSGDF 20 98 YTSLWDLRHL 20 353 EVWRIEMYIS 20 68 EFFPHVVDVT 19 75 DVTHHEDALT 19 115 DVSNNMRINQ 19 186 NFIPIDLGSL 19 230 SFVRDVIHPY 19 257 KTLPIVAITL 19 314 MVHVAYSLCL 19 364 GIMSLGLLSL 19 404 LISTFHVLIY 19 217 VVAISLATFF 18 359 MYISFGIMSL 18 399 GYVALLISTF 18 441 IVILDLLQLC 18 2 ESISMMGSPK 17 30 KVTVGVIGSG 17 40 DFAKSLTIRL 17 81 DALTKTNIIF 17 263 AITLLSLVYL 17 406 STFHVLIYGW 17 177 QVIELARQLN 16 215 PVVVAISLAT 16 269 LVYLAGLLAA 16 435 ALVLPSIVIL 16 436 LVLPSIVILD 16 34 GVIGSGDFAK 15 72 HVVDVTHHED 15 116 VSNNMRINQY 15 142 IVKGFNVVSA 15 199 REIENLPLRL 15 250 IPIEIVNKTL 15 261 IVAITLLSLV 15 262 VAITLLSLVY 15 310 FFFAMVHVAY 15 377 TSIPSVSNAL 15 389 REFSFIQSTL 15 391 FSFIQSTLGY 15 432 FVLALVLPSI 15 31 VTVGVIGSGD 14 55 HVVIGSRNPK 14 89 IFVAIHREHY 14 103 DLRHLLVGKI 14 108 LVGKILIDVS 14 148 VVSAWALQLG 14 222 LATFFFLYSF 14 301 CRKQLGLLSF 14 352 EEVWRIEMYI 14 362 SFGIMSLGLL 14 417 RAFEEEYYRF 14 437 VLPSIVILDL 14 443 ILDLLQLCRY 14 27 DARKVTVGVI 13 74 VDVTHHEDAL 13 92 AIHREHYTSL 13 137 FPDSLIVKGF 13 172 IQARQQVIEL 13 176 QQVIELARQL 13 178 VIELARQLNF 13 218 VAISLATFFF 13 223 ATFFFLYSFV 13 258 TLPIVAITLL 13 299 LQCRKQLGLL 13 302 RKQLGLLSFF 13 358 EMYISFGIMS 13 361 ISFGIMSLGL 13 365 IMSLGLLSLL 13 375 AVTSIPSVSN 13 376 VTSIPSVSNA 13 395 QSTLGYVALL 13 410 VLIYGWKRAF 13

TABLE XXXVIII-V2 HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 17 ETPFSCLSLP 13 16 GFTPFSGLSL 12 35 PPCPADFFLY 11 2 GSPGLQALSL 10 4 PGLQALSLSL 10 14 SSGFTPFSCL 10 22 CLSLPSSWDY 10 8 ALSLSLSSGF 9 11 LSLSSGFTPF 9 32 RCPPPCPADF 9 33 CPPPCPADFF 9 36 PCPADFFLYF 9 30 DYRCPPPCPA 8 34 PPPCPADFFL 8 7 QALSLSLSSG 7 18 TPFSCLSLPS 7 3 SPGLQALSLS 6

TABLE XXXVIII-V5A HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 1 ENLPLRLFTF 24 8 FTFWRGPVVV 12

TABLE XXXVIII-V5B HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 16 DTQTELELEF 25 22 ELEFVFLLTL 24 24 EFVFLLTLLL 23 20 ELELEFVFLL 22 18 QTELELEFVF 16 23 LEFVFLLTLL 16 4 EFSFIQIFCS 14 5 FSFIQIFCSF 13 2 WREFSFIQIF 12 12 GSFADTQTEL 12

TAABLE XXXVIII-V6 HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 6 IVILGKILLF 27 5 SIVILGKIIL 18 38 EGIGGTIPHV 18 7 VILGKIILFL 17 1 LVLPSTVILG 16 46 HVSPERVTVM 15 42 GTLPHVSPER 13

TABLE XXXVIII-V7A HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 ETFLPNGING 24

TABLE XXXVIII-V7B HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 9 QSTLGYVALL 13 5 MAYQQSTLGY 11 3 LNMAYQQSTL 10 10 STLGYVALLI 10 8 QQSTLGYVAL 9

TABLE XXXVIII-V7C HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 170 ETIILSKLTQ 24 12 EVLASPAAAW 21 35 EIVLPIEWQQ 19 102 DPPESPDRAL 19 127 GVGPLWEFLL 19 5 VILDLSVEVL 17 152 FTSWSLGEFL 17 69 EAQESGIENK 16 105 ESPDRALKAA 16 89 GVVTEDDEAQ 15 133 EFLLRLLKSQ 15 151 AFTSWSLGEF 15 3 SIVILDLSVE 14 4 IVILDLSVEV 14 45 DRKIPPLSTP 14 86 PVVGVVTEDD 14 90 VVTEDDEAQD 14 99 DSIDPPESPD 14 130 PLWEFLLRLL 14 168 KLETIILSKL 14 171 TIILSKLTQE 14 8 DLSVEVLASP 13 42 WQQDRKIPPL 13 93 EDDEAQDSID 13 122 LPHTNGVGPL 13 125 TNGVGPLWEF 13 129 GPLWEFLLRL 13 10 SVEVLASPAA 12 36 IVLPIEWQQD 12 72 ESGIRNKSSS 12 95 DEAQDSIDPP 12 120 PVLPHTNGVG 12 126 NGVGPLWEFL 12 41 EWQQDRKIPP 11 60 WTEEAGATAE 11 62 EEAGATAEAQ 11 63 EAGATAEAQE 11 66 ATAEAQESGI 11 96 EAQDSIDPPE 11 141 SQAASGTLSL 11 159 EFLGSGTWMK 11 180 EQKSKHCMFS 11

TABLE XXXVIII-V8 HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 EGMGGTIPHV 14 7 EEGMGGTIPH 11 1 EKSQFLEEGM 10 3 SQFLEEGMGG 6

TABLE XXXVIII-V13 HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 ETFLPNGING 24

TABLE XXXVIII-V14 HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 1 ENLPLRLFTF 24 8 FTFWRGPVVV 12

TABLE XXXVIII-V21 HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 4 LTQEQKTKHG 10 7 EQKTKHCMFS 10 8 QKTKHCMFSL 10 6 QEQKTKHCMF 9 9 KTKHCMFSLI 9

TABLE XXXVIII-V25 HLA-A26-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 2 ILFLPGISQK 10 3 LFLPCISQKL 10 6 PCISQKLKRI 9 1 IILFLPCISQ 6 9 SQKLKRIKKG 6 7 CISQKLKRIK 4

TABLE XXXIX-V1 HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 429 PPNFVLALVL 23 438 LPS1VILDLL 22 9 SPKSLSETCL 21 250 IPIEIVNKTL 21 323 LPMRRSERYL 21 137 FPDSLIVKGF 18 428 TPPNFVLALV 17 125 YPESNAEYLA 16 214 GPVVVAISLA 16 219 AISLATFFFL 16 394 IQSTLGYVAL 16 36 IGSGDFAKSL 15 197 SAREIENLPL 15 325 MRRSERYLFL 15 361 ISFGIMSLGL 15 379 IPSVSNALNW 15 427 YTPPNFVLAL 15 211 LWRGPVVVAI 14 263 AITLLSLVYL 14 402 ALLISTFHVL 14 435 ALVLPSIVIL 14 40 DFAKSLTIRL 13 92 AIHREHYTSL 13 127 ESNAEYLASL 13 172 IQARQQVIEL 13 188 IPIDLGSLSS 13 195 LSSAREIENL 13 199 REIENLPLRL 13 204 LPLRLFTLWR 13 259 LPIVAITLLS 13 260 PIVAITLLSL 13 266 LLSLVYLAGL 13 290 REPPWLETWL 13 364 GIMSLGLLSL 13 365 IMSLGLLSLL 13 4 ISMMGSPKSL 12 18 LPNGINGIKD 12 70 FPHVVDVTHH 12 98 YTSLWDLRHL 12 142 IVKGFNVVSA 12 147 NVVSAWALQL 12 157 GPKDASRQVY 12 202 ENLPLRLFTL 12 257 KTLPIVAITL 12 273 AGLLAAAYQL 12 292 PPWLETWLQC 12 296 ETWLQCRKQL 12 298 WLQCRKQLGL 12 314 MVHVAYSLCL 12 377 TSIPSVSNAL 12 395 QSTLGYVALL 12 425 RFYTPPNFVL 12 437 VLPSIVILDL 12 440 SIVILDLLQL 12 26 KDARKVTVGV 11 27 DARKVTVGVI 11 49 LIRCGYHVVI 11 62 NPKFASEFFP 11 74 VDVTHHEDAL 11 95 REHYTSLWDL 11 99 TSLWDLRHLL 11 132 YLASLFPDSL 11 145 GFNVVSAWAL 11 183 RQLNFIPIDL 11 186 NFIPIDLGSL 11 201 IENLPLRLFT 11 213 RGPVVVAISL 11 237 HPYARNQQSD 11 252 IEIVNKTLPI 11 258 TLPIVAITLL 11 286 TKYRRFPPWL 11 291 FPPWLETWLQ 11 312 FAMVHVAYSL 11 362 SFGIMSLGLL 11 389 REFSFIQSTL 11

TABLE XXXIX-V2 HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 34 PPPCPADFFL 21 33 CPPPCPADFF 18 2 GSPGLQALSL 14 16 GFTPFSCLSL 13 18 TPFSCLSLPS 13 4 PGLQALSLSL 12 14 SSGFTPFSCL 12 25 LPSSWDYRCP 12 35 PPGPADFFLY 12 3 SPGLQALSLS 11 8 ALSLSLSSGF 10 36 PGPADFFLYF 10

TABLE XXXIX-V5A HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 10 FWRGPVVVAI 14 3 LPLRLFTFWR 11 9 TFWRGPVVVA 10 6 RLFTFWRGPV 9 8 FTFWRGPVVV 9 1 ENLPLRLFTF 8 7 LFTFWRGPVV 8

TABLE XXXIX-V5B HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 19 TELELEFVFL 14 24 EFVFLLTLLL 14 14 FADTQTELEL 13 22 ELEFVFLLTL 13 12 CSFADTQTEL 12 20 ELELEFVFLL 12 23 LEFVFLLTLL 11 1 NWREFSFIQI 9 8 IQIFCSFADT 9 21 LELEFVFLLT 9 10 IFCSFADTQT 8 16 DTQTELELEF 8 5 FSFIQIFGSF 7 6 SFIQIFCSFA 7 17 TQTELELEFV 7 18 QTELELEFVF 7 2 WREFSFIQIF 6

TABLE XXXIX-V6 HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 3 LPSIVILGKI 18 44 IPHVSPERVT 18 7 VILGKIILFL 15 27 KKGWEKSQFL 13 16 LPCISRKLKR 12 46 HVSPERVTVM 12 14 LFLPCISRKL 11 5 SIVILGKIIL 10 38 EGIGGTIPHV 10 26 IKKGWEKSQF 9 31 EKSQFLEEGI 9 45 PHVSPERVTV 9

TABLE XXXIX-V7A HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 2 SPKSLSETFL 22

TABLE XXXIX-V7B HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 QQSTLGYVAL 15 3 LNMAYQQSTL 12 9 QSTLGYVALL 12 10 STLGYVALLI 10 6 AYQQSTLGYV 8 7 YQQSTLGYVA 7

TABLE XXXIX-V7C HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 122 LPHTNGVGPL 22 129 GPLWEFLLRL 22 102 DPPESPDRAL 21 49 PPLSTPPPPA 18 55 PPPAMWTEEA 18 119 NPVLPHTNGV 17 141 SQAASGTLSL 15 143 AASGTLSLAF 15 29 LRGGLSEIVL 14 113 AANSWRNPVL 14 15 ASPAAAWKCL 13 48 IPPLSTPPPP 13 85 IPVVGVVTED 13 106 SPDRALKAAN 13 126 NGVGPLWEFL 13 152 FTSWSLGEFL 13 165 TWMKLETIIL 13 181 QKSKHCMFSL 13 1 LPSIVILDLS 12 5 VILDLSVEVL 12 16 SPAAAWKCLG 12 20 AWKCLGANIL 12 24 LGANILRGGL 12 42 WQQDRKIPPL 12 54 PPPPAMWTEE 12 56 PPAMWTEEAG 12 103 PPESPDRALK 12 127 GVGPLWEFLL 12 139 LKSQAASGTL 12 28 ILRGGLSEIV 11 44 QDRKIPPLST 11 53 TPPPPAMWTE 11 81 SSSQIPVVGV 11 104 PESPDRALKA 11 144 ASGTLSLAFT 11 148 LSLAFTSWSL 11 160 FLGSGTWMKL 11 168 KLETIILSKL 11 6 ILDLSVEVLA 10 17 PAAAWKCLGA 10 19 AAWKCLGANI 10 31 GGLSEIVLPI 10 38 LPIEWQQDRK 10 50 PLSTPPPPAM 10 78 KSSSSSQIPV 10 79 SSSSSQIPVV 10 83 SQIPVVGVVT 10 112 KAANSWRNPV 10 130 PLWEFLLRLL 10

TABLE XXXIX-V8 HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 EGMGGTIPHV 11 1 EKSQFLEEGM 9 4 QFLEEGMGGT 6 5 FLEEGMGGTI 6

TABLE XXXIX-V13 HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 2 SPKSLSETFL 22

TABLE XXXIX-V14 HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 10 FWRGPVVVAI 14 3 LPLRLFTFWR 11 9 TFWRGPVVVA 10 6 RLFTFWRGPV 9 8 FTFWRGPVVV 9 1 ENLPLRLFTF 8 7 LFTFWRGPVV 8

TABLE XXXIX-V21 HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 QKTKHCMFSL 11 9 KTKHCMFSLI 8 6 QEQKTKHCMF 7 1 LSKLTQEQKT 6 5 TQEQKTKHGM 6

TABLE XXXIX-V25 HLA-B0702-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 5 LPCISQKLKR 12 3 LFLPCISQKL 11 6 PCISQKLKRI 6

TABLE XL-V1 HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V2 HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V5A HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V5B HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V6 HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V7A HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V7B HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V7C HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V8 HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V13 HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V14 HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V21 HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XL-V25 HLA-B08-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V1 HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V2 HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V5A HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V5B HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V6 HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V7A HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V7B HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V7C HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V8 HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V13 HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V14 HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V21 HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V25 HLA-B1510-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V1 HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V2 HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V5A HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V5B HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V6 HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V7A HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V7B HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V7C HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V8 HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V13 HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V14 HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V21 HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V25 HLA-B2705-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V1 HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V2 HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V5A HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V5B HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V6- HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V7A HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V7B HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V7C HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V8 HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V13 HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V14 HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V21 HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLI-V25 HLA-B2709-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLIV-V1 HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 199 REIENLPLRL 25 351 EEEVWRIEMY 25 252 IEIVNKTLPI 23 389 REFSFIQSTL 23 95 REHYTSLWDL 21 179 IELARQLNFI 21 352 EEVWRIEMYI 20 79 HEDALTKTNI 19 377 TSIPSVSNAL 19 186 NFIPIDLGSL 18 202 ENLPLRLFTL 18 257 KTLPIVAITL 18 427 YTPPNFVLAL 18 435 ALVLPSIYIL 18 273 AGLLAAAYQL 17 289 RRFPPWLETW 17 296 ETWLQGRKQL 17 402 ALLISTFHVL 17 16 TCLPNGINGI 16 116 VSNNMRINQY 16 200 EIENLPLRLF 16 219 AISLATFFFL 16 230 SFVRDVIHPY 16 250 IPIETVNKTL 16 262 VAITLLSLVY 16 263 AITLLSLVYL 16 359 MYISFGTMSL 16 406 STFHVLIYGW 16 410 VLIYGWKRAF 16 36 IGSGDFAKSL 15 45 LTIRLIRCGY 15 56 VVIGSRNPKF 15 60 SRNPKFASEF 15 67 SEFFPHVVDV 15 126 PESNAEYLAS 15 130 AEYLASLFPD 15 203 NLPLRLFTLW 15 255 VNKTLPIVAI 15 258 TLPIVAITLL 15 279 AYQLYYGTKY 15 310 FFFAMVHVAY 15 329 ERYLFLNMAY 15 394 IQSTLGYVAL 15 437 VLPSIVILDL 15 4 ISMMGSPKSL 14 92 AIHREHYTSL 14 98 YTSLWDLRHL 14 99 TSLWDLRHLL 14 123 NQYPESNAEY 14 137 FPDSLIVKGF 14 147 NVVSAWALQL 14 183 RQLNFIPIDL 14 195 LSSAREIENL 14 218 VAISLATFFF 14 271 YLAGLLAAAY 14 290 RFPPWLETWL 14 346 ENSWNEEEVW 14 361 ISFGIMSLGL 14 365 IMSLGLLSLL 14 391 FSFIQSTLGY 14 396 STLGYVALLI 14 399 GYVALLISTF 14 404 LISTFHVLIY 14 418 AFEEEYYRFY 14 420 EEEYYRFYTP 14 440 SIVILDLLQL 14 41 FAKSLTIRLI 13 74 VDVTHHEDAL 13 80 EDALTKTNII 13 81 DALTKTNIIF 13 84 TKTNIIFVAI 13 104 LRHLLVGKIL 13 127 ESNAEYLASL 13 128 SNAEYLASLF 13 143 VKGFNVVSAW 13 145 GFNVVSAWAL 13 157 GPKDASRQVY 13 170 NNIQARQQVI 13 172 IQARQQVIEL 13 176 QQVIELARQL 13 201 IENLPLRLFT 13 211 LWRGPVVVAI 13 213 RGPVVVAISL 13 220 ISLATFFFLY 13 245 SDFYKIPIEI 13 266 LLSLVYLAGL 13 267 LSLVYLAGLL 13 299 LQGRKQLGLL 13 303 KQLGLLSFFF 13 323 LPMRRSERYL 13 324 PMRRSERYLF 13 328 SERYLFLNMA 13 350 NEEEVWRIEM 13 362 SFGIMSLGLL 13 364 GIMSLGLLSL 13 379 IPSVSNALNW 13 384 NALNWREFSF 13 395 QSTLGYVALL 13 403 LLISTFHVLI 13 429 PPNFVLALVL 13 438 LPSIVILDLL 13 443 ILDLLQLCRY 13 38 SGDFAKSLTI 12 40 DFAKSLTIRL 12 93 IHREHYTSLW 12 105 RHLLVGKILI 12 124 QYPESNAEYL 12 178 VIELARQLNF 12 192 LGSLSSAREI 12 197 SAREIENLPL 12 216 VVVAISLATF 12 260 PIVAITLLSL 12 274 GLLAAAYQLY 12 282 LYYGTKYRRF 12 286 TKYRRFPPWL 12 295 LETWLQCRKQ 12 301 CRKQLGLLSF 12 302 RKQLGLLSFF 12 312 FAMVHVAYSL 12 357 IEMYISFGIM 12 385 ALNWREFSFI 12 417 RAFEEEYYRF 12 421 EEYYRFYTPP 12 425 RFYTPPNFVL 12

TABLE XLIV-V2 HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 ALSLSLSSGF 15 32 RCPPPCPADF 15 33 CPPPCPADFF 15 35 PPCPADFFLY 15 2 GSPGLQALSL 14 16 GFTPFSCLSL 14 36 PCPADFFLYF 13 4 PGLQALSLSL 12 11 LSLSSGFTPF 12 14 SSGFTPFSCL 12 20 FSCLSLPSSW 12 22 CLSLPSSWDY 12 34 PPPCPADFFL 11

TABLE XLIV-V5A HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 1 ENLPLRLFTF 18 2 NLPLRLFTFW 14 10 FWRGPVVVAI 13

TABLE XLIV-V5B HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 23 LEFVFLLTLL 24 19 TELELEFVFL 23 20 ELELEFVFLL 15 22 ELEFVFLLTL 15 24 EFVFLLTLLL 15 21 LELEFVELLT 14 2 WREFSFIQIF 13 3 REFSFIQIFC 13 5 FSFIQIFCSF 13 14 FADTQTELEL 13 1 NWREFSFIQI 12 12 GSFADTQTEL 12 16 DTQTELELEF 12 18 QTELELEFVF 12

TABLE XLIV-V6 HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 6 IVILGKIILF 19 7 VLLGKIILFL 16 14 LFLPCISRKL 16 17 PCISRKLKRI 14 37 EEGIGGTIPH 14 4 PSIVILGKII 13 21 RKLKRJKKGW 13 5 SIVILGKIIL 12 10 GKIILFLPCI 12 26 IKKGWEKSQF 12 3 LPSIVILGKI 11 27 KKGWEKSQFL 11 30 WEKSQFLEEG 11 31 EKSQFLEEGI 11 36 LEEGIGGTIP 11 35 FLEEGIGGTI 9 38 EGIGGTIPHV 9

TABLE XLIV-V7A HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 9 TFLPNGINGI 16 1 GSPKSLSETF 12 2 SPKSLSETFL 11 6 LSETFLPNGI 11 7 SETFLPNGIN 11

TABLE XLIV-V7B HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 8 QQSTLGYVAL 15 10 STLGYVALLI 14 9 QSTLGYVALL 13 3 LNMAYQQSTL 12 5 MAYQQSTLGY 12

TABLE XLIV-V7C HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 92 TEDDEAQDSI 20 179 QEQKSKHCMF 20 143 AASGTLSLAF 18 34 SEIVLPIEWQ 17 104 PESPDRALKA 17 12 EVLASPAAAW 16 15 ASPAAAWKCL 16 62 EEAGATAEAQ 16 132 WEFLLRLLKS 16 20 AWKCLGANIL 15 5 VILDLSVEVL 14 11 VEVLASPAAA 14 42 WQQDRKIPPL 14 51 LSTPPPPAMW 14 68 AEAQESGIRN 14 71 QESGIRNKSS 14 102 DPPESPDRAL 14 113 AANSWRNPVL 14 127 GVGPLWEFLL 14 151 AFTSWSLGEF 14 168 KLETIILSKL 14 29 LRGGLSEIVL 13 40 IEWQQDRKIP 13 95 DEAQDSIDPP 13 108 DRALKAANSW 13 129 GPLWEFLLRL 13 130 PLWEFLLRLL 13 141 SQAASGTLSL 13 158 GEFLGSGTWM 13 165 TWMKLETIIL 13 169 LETIILSKLT 13 24 LGANILRGGL 12 27 NILRGGLSEI 12 33 LSEIVLPIEW 12 122 LPHTNGVGPL 12 123 PHTNGVGPLW 12 126 NGVGPLWEFL 12 139 LKSQAASGTL 12 146 GTLSLAFTSW 12 19 AAWKGLGAN7 11 31 GGLSEIVLPI 11 61 TEEAGATAEA 11 66 ATAEAQESGI 11 125 TNGVGPLWEF 11 148 LSLAFTSWSL 11 152 FTSWSLGEFL 11 157 LGEFLGSGTW 11 160 FLGSGTWMKI 11 163 SGTWMKLETI 11 181 QKSKHCMFSL 11 182 KSKIHCMFSL 11 39 PIEWQQDRKI 10 76 RNLSSSSSQI 9 83 SQIPVVGVVT 9 105 ESPDRALKAA 9

TABLE XLIV-V8 HLA-B4402-10Omers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 7 EEGMGGTIPH 14 6 LEEGMGGTIP 11 5 FLEEGMGGTI 9 8 EGMGGTIPHV 7

TABLE XLIV-V13 HLA-B4402-10Omers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 9 TFLPNGINGI 16 1 GSPKSLSETF 12 2 SPKSLSETFL 11 6 LSETFLPNGI 11 7 SETFLPNGIN 11

TABLE XLIV-V14 HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 1 ENLPLRLFTF 18 2 NLPLRLFTFW 14 10 FWRGPVVVAI 13

TABLE XLIV-V21 HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 6 QEQKTLHCMF 20 9 KTKHCMFSLI 11 8 QKTKHCMFSL 10

TABLE XLIV-V25 HLA-B4402-10mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. Pos 1234567890 score 3 LFLPCISQKL 15 6 PCISQKLKRI 14 10 QKLKRLKKGW 13 9 SQKLKRIKKG 8 2 ILFLPCISQK 7

TABLE XLV-V1 HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V2 HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V5A HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V5B HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V6 HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V7A HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V7B HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V7C HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V8 HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V13 HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V14 HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V21 HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLV-V25 HLA-B5101-10mers-98P4B6 Pos 1234567890 score NoResultsFound.

TABLE XLVI-V1 HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 143 VKGFNVVSAWALQLG 33 266 LLSLVYLAGLLAAAY 33 367 SLGLLSLLAVTSIPS 32 1 MESISMMGSPKSLSE 31 130 AEYLASLFPDSLIVK 30 30 KVTVGVIGSGDFAKS 29 431 NFVLALVLPSIVILD 29 206 LRLFTLWRGPVVVAI 28 215 PVVVAISLATFFFLY 28 370 LLSLLAVTSIPSVSN 28 438 LPSIVILDLLQLCRY 28 101 LWDLRHLLVGKILID 27 185 LNFIPIDLGSLSSAR 27 356 RIEMYISFGIMSLGL 27 360 YISFGIMSLGLLSLL 27 397 TLGYVALLISTFHVL 27 421 EEYYRFYTPPNFVLA 27 38 SGDFAKSLTIRLIRC 26 102 WDLRHLLVGKILIDV 26 122 INQYPESNAEYLASL 26 149 VSAWALQLGPKDASR 26 244 QSDFYKIPIEIVNKT 26 249 KIPIEIVNKTLPIVA 26 256 NKTLPIVAITLLSLV 26 261 IVAITLLSLVYLAGL 26 298 WLQCRKQLGLLSFFF 26 368 LGLLSLLAVTSIPSV 26 109 VGKILIDVSNNMRIN 25 137 FPDSLIVKGFNVVSA 25 145 GFNVVSAWALQLGPK 25 198 AREIENLPLRLFTLW 25 222 LATFFFLYSFVRDVI 25 252 EIEVNKTLPIVAITL 25 264 ITLLSLVYLAGLLAA 25 302 RKQLGLLSFFFAMVH 25 309 SFFFAMVHVAYSLCL 25 354 VWRIEMYISFGIMSL 25 362 SFGIMSLGLLSLLAV 25 365 IMSLGLLSLLAVTSI 25 51 RCGYHVVIGSRNPKF 24 98 YTSLWDLRHLLVGKI 24 106 HLLVGKILIDVSNNM 24 150 SAWALQLGPKDASRQ 24 184 QLNFIPIDLGSLSSA 24 205 PLRLFTLWRGPVVVA 24 229 YSFVRDVIHPYARNQ 24 269 LVYLAGLLAAAYQLY 24 330 RYLFLNMAYQQVHAN 24 335 NMAYQQVHANIENSW 24 388 WREFSFIQSTLGYVA 24 391 FSFIQSTLGYVALLI 24 398 LGYVALLISTFHVLI 24 427 YTPPNFVLALVLPSI 24 430 PNFVLALVLPSIVIL 24 52 CGYHVVIGSRNPKFA 23 55 HVVIGSRNPKFASEF 23 186 NFIPIDLGSLSSARE 23 214 GPVVVAISLATFFFL 23 258 TLPIVAITLLSLVYL 23 351 EEEVWRIEMYISFGI 23 352 EEVWRIEMYISFGIM 23 127 ESNAEYLASLFPDSL 22 178 VIELARQLNFIPIDL 22 189 PIDLGSLSSAREIEN 22 211 LWRGPVVVAISLATF 22 216 VVVAISLATFFFLYS 22 255 VNKTLPIVAITLLSL 22 301 CRKQLGLLSFFFAMV 22 312 FAMVHVAYSLCLPMR 22 359 MYISFGIMSLGLLSL 22 364 GIMSLGLLSLLAVTS 22 395 QSTLGYVALLISTFH 22 432 FVLALVLPSIVILDL 22 435 ALVLPSIVILDLLQL 22 20 NGINGIKDARKVTVG 21 117 SNNMRINQYPESNAE 21 161 ASRQVYICSNNIQAR 21 174 ARQQVIELARQLNFI 21 277 AAAYQLYYGTKYRRF 21 373 LLAVTSIPSVSNALN 21 399 GYVALLISTFHVLIY 21 407 TFHVLIYGWKRAFEE 21 31 VTVGVIGSGDFAKSL 20 142 IVKGFNVVSAWALQL 20 209 FTLWRGPVVVAISLA 20 346 ENSWNEEEVWRIEMY 20 385 ALNWREFSFIQSTLG 20 429 PPNFVLALVLPSIVI 20 45 LTIRLIRCGYHVVIG 19 80 EDALTKTNIIFVAIH 19 95 REHYTSLWDLRHLLV 19 135 SLFPDSLIVKGFNVV 19 139 DSLIVKGFNVVSAWA 19 224 TFFFLYSFVRDVIHP 19 259 LPIVAITLLSLVYLA 19 280 YQLYYGTKYRRFPPW 19 281 QLYYGTKYRRFPPWL 19 288 YRRFPPWLETWLQCR 19 307 LLSFFFAMVHVAYSL 19 322 CLPMRRSERYLFLNM 19 328 SERYLFLNMAYQQVH 19 357 IEMYISFGIMSLGLL 19 400 YVALLISTFHVLIYG 19 424 YRFYTPPNFVLALVL 19 7 MGSPKSLSETCLPNG 18 25 IKDARKVTVGVIGSG 18 27 DARKVTVGVIGSGDF 18 39 GDFAKSLTIRLIRCG 18 47 IRLIRCGYHVVIGSR 18 62 NPKFASEFFPHVVDV 18 129 NAEYLASLFPDSLIV 18 163 RQVYICSNNIQARQQ 18 167 ICSNNIQARQQVIEL 18 179 IELARQLNFIPIDLG 18 190 IDLGSLSSAREIENL 18 236 IHPYARNQQSDFYKI 18 267 LSLVYLAGLLAAAYQ 18 268 SLVYLAGLLAAAYQL 18 285 GTKYRRFFPWLETWL 18 296 ETWLQCRKQLGLLSF 18 299 LQCRKQLGLLSFFFA 18 326 RRSERYLFLNMAYQQ 18 380 PSVSNALNWREFSFI 18 383 SNALNWREFSFIQST 18 390 EFSFIQSTLGYVALL 18 405 ISTFHVLIYGWKRAF 18 410 VLIYGWKRAFEEEYY 18 423 YYRFYTPPNFVLALV 18 433 VLALVLPSLVILDLL 18 22 INGIKDARKVTVGVI 17 29 RKVTVGVIGSGDFAK 17 33 VGVIGSGDFAKSLTI 17 34 GVIGSGDFAKSLTIR 17 44 SLTIRLIRCGYHVVI 17 46 TIRLIRGGYHVVIGS 17 54 YHVVIGSRNPKFASE 17 58 IGSRNPKFASEFFPH 17 77 THHEDALTKTNIIFV 17 87 NIIFVAIHREHYTSL 17 90 FVAIHREHYTSLWDL 17 105 RHLLVGKILIDVSNN 17 119 NMRINQYPESNAEYL 17 138 PDSLIVKGFNVVSAW 17 140 SLIVKGFNVVSAWAL 17 151 AWALQLGPKDASRQV 17 154 LQLGPKDASRQVYIC 17 176 QQVIELARQLNFIPI 17 187 FIPIDLGSLSSAREI 17 195 LSSAREIENLPLRLF 17 217 VVAISLATFFFLYSF 17 226 FFLYSFVRDVIHPYA 17 232 VRDVIHPYARNQQSD 17 251 PIEIVNKTLPIVAIT 17 253 EIVNKTLPIVAITLL 17 270 VYLAGLLAAAYQLYY 17 271 YLAGLLAAAYQLYYG 17 305 LGLLSFFFAMVHVAY 17 316 HVAYSLCLPMRRSER 17 317 VAYSLCLPMRRSERY 17 329 ERYLFLNMAYQQVHA 17 361 ISFGIMSLGLLSLLA 17 363 FGIMSLGLLSLLAVT 17 389 REFSFIQSTLGYVAL 17 392 SFIQSTLGYVALLIS 17 406 STFHVLIYGWKRAFE 17 408 FHVLIYGWKRAFEEE 17 436 LVLPSIVILDLLQLC 17 2 ESISMMGSPKSLSET 16 3 SISMMGSPKSLSETC 16 8 GSPKSLSETCLPNGI 16 11 KSLSETGLPNGINGI 16 16 TCLPNG1NGIKDARK 16 24 GIKDARKVTVGVIGS 16 59 GSRNPKFASEFFPHV 16 67 SEFFPHVVDVTHHED 16 71 PHVVDVTHHEDALTK 16 103 DLRHLLVGKILIDVS 16 111 KILIDVSNNMRINQY 16 126 PESNAEYLASLFPDS 16 153 ALQLGPKDASRQVYI 16 166 YICSNNIQARQQVIE 16 171 NIQARQQVIELARQL 16 175 RQQVIELARQLNFIP 16 182 ARQLNFIPIDLGSLS 16 200 EIENLPLRLFTLWRG 16 208 LFTLWRGPVVVAISL 16 219 AISLATFFFLYSFVR 16 225 FFFLYSFVRDVIHPY 16 263 AITLISLVYLAGLLA 16 265 TLLSLVYLAGLLAAA 16 294 QLETWLQCRKQLGLL 16 304 QLGLLSFFFAMVHVA 16 308 LSFFFAMVHVAYSLC 16 310 FFFAMVHVAYSLCLP 16 314 MVHVAYSLCLPMRRS 16 371 LSLLAVTSIPSVSNA 16 394 IQSTLGYVALLISTF 16 401 VALLISTFHVLIYGW 16 420 EEEYYRFYTPPNFVL 16 428 TPPNFVLALVLPSIV 16 440 SIVILDLLQLCRYPD 16

TABLE XLVI-V2 HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 17 FTPFSCLSLPSSWDY 26 28 SWDYRGPPPCPADFF 26 6 LQALSLSLSSGFTPF 25 8 ALSLSLSSGFTPFSC 25 3 SPGLQALSLSLSSGF 24 10 SLSLSSGFTPFSCLS 22 14 SSGFTPFSCLSLPSS 19 26 PSSWDYRCPPPCPAD 16 31 YRCPPPCPADFFLYF 16 1 SGSPGLQALSLSLSS 15 4 PGLQALSLSLSSGFT 15 20 FSCLSLPSSWDYRCP 15 2 GSPGLQALSLSLSSG 14 7 QALSLSLSSGFTPFS 14 13 LSSGFTPFSCLSLPS 14 16 GFTPFSGLSLPSSWD 14 19 PFSGLSLPSSWDYRC 14 27 SSWDYRGPPPGPADF 14 30 DYRCPPPCPADFFLY 14

TABLE XLVI-V5A HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 11 LRLFTFWRGPVVVAI 28 3 AREIENLPLRLFTFW 25 16 FWRGPVVVAISLATF 22 14 FTFWRGPVVVAISLA 20 13 LFTFWRGPVVVAISL 18 5 EIENLPLRLFTFWRG 16 10 PLRLFTFWRGPVVVA 16 12 RLFTFWRGPVVVAIS 15 2 SAREIENLPLRLFTF 14 7 ENLPLRLFTFWRGPV 14 15 TFWRGPVVVAISLAT 14

TABLE XLVI-V5B HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 7 WREFSFIQIFGSFAD 25 9 EFSFIQIEGSFADTQ 24 4 ALNWREFSFIQIFGS 20 2 SNALNWREFSFIQIF 18 20 ADTQTELELEFVFLL 18 8 REFSFIQIFCSFADT 17 10 FSFIQIFGSFADTQT 17 22 TQTELELEFVFLLTL 17 23 QTELELEFVFLLTLL 17 12 FIQIFCSFADTQTEL 16 16 FCSFADTQTELELEF 16 17 CSFADTQTELELEFV 14

TABLE XLVI-V6 HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 1 NFVLALVLPSIVILG 29 8 LPSIVILGKIILFLP 29 46 GGTIPHVSPERVTVM 28 17 IILFLPCISRKLKRI 26 11 IVILGKIILFLPCIS 24 38 SQFLEEGIGGTIPHV 24 39 QFLEEGIGGTIPHVS 24 7 VLPSIVLLGKIILFL 23 14 LGKIILFLPCISRKL 23 2 FVLALVLPSIVILGK 22 42 EEGIGGTIPHVSPER 22 13 ILGKIILFLPCISRK 19 3 VLALVLPSIVILGKI 18 6 LVLPSIVILGKflLF 18 9 PSIVILGKIILFLPG 17 15 GKIILFLPCISRKLK 17 5 ALVLPSIVILGKIIL 16 10 SIVILGKIILFLPCI 16 18 ILFLPCISRKLKRIK 15 25 SRKLKRIKKGWEKSQ 15 30 RIKKGWEKSQFLEEG 14 43 EGIGGTIPHVSPERV 14

TABLE XLVI-V7A HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 12 SETFLPNGINGIKDA 21 5 MGSPKSLSETFLPNG 18 1 SISMMGSPKSLSETF 16 4 MMGSPKSLSETFLPN 16 6 GSPKSLSETFLPNGI 16 9 KSLSETFLPNGINGI 16 14 TFLPNGINGIKDARK 16 2 ISMMGSPKSLSETFL 14 15 FLPNG1NGIKDARKV 13 10 SLSETFLPNGINGIK 10

TABLE XLVI-V7B HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 4 RYLFLNMAYQQSTLG 24 14 QSTLGYVALLISTFH 22 7 FLNMAYQQSTLGYVA 21 2 SERYLFLNMAYQQST 19 9 NMAYQQSTLGYVALL 18 3 ERYLFLNMAYQQSTL 17 11 AYQQSTLGYVALLIS 17 10 MAYQQSTLGYVALLI 16 13 QQSTLGYVALLISTF 16 8 LNMAYQQSTLGYVAL 14

TABLE XLVI-V7C HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 23 AAAWKCLGANILRGG 36 168 SGTWMKLETIILSKL 35 138 EFLLRLLKSQAASGT 33 13 DLSVEVLASPAAAWK 30 50 DRKIPPLSTPPPPAM 30 28 CLGANILRGGLSEIV 28 62 PAMWTEEAGATAEAQ 27 110 ESPDRALKAANSWRN 26 124 NPVLPHTNGVGPLWE 26 141 LRLLKSQAASGTLSL 25 8 SIVILDLSVEVLASP 24 31 ANILRGGLSEIVLPI 24 42 VLPIEWQQDRKIPPL 24 77 ESGIRNKSSSSSQIP 24 130 TNGVGPLWEFLLRLL 24 137 WEFLLRLLKSQAASG 24 7 PSIVILDLSVEVLAS 23 12 LDLSVEVLASPAAAW 23 150 SGTLSLAFTSWSLGE 23 171 WMKLETIILSKLTQE 23 3 ALVLPSIVILDLSVE 22 53 IPPLSTPPPPAMWTE 22 157 FTSWSLGEFLGSGTW 22 89 QIPVVGVVTEDDEAQ 21 6 LPSIVILDLSVEVLA 20 58 TPPPPAMWTEEAGAT 20 97 TEDDEAQDSIDPPES 20 100 DEAQDSIDPPESPDR 20 134 GPLWEFLLRLLKSQA 19 154 SLAFTSWSLGEFLGS 19 1 VLALVLPSIVILDLS 18 22 PAAAWKCLGANILRG 18 44 PIEWQQDRKIPPLST 18 122 WRNPVLPHTNGVGPL 18 135 PLWEFLLRLLKSQAA 18 140 LLRLLKSQAASGTLS 18 148 AASGTLSLAFTSWSL 18 159 SWSLGEFLGSGTWMK 18 161 SLGEFLGSGTWMKLE 18 169 GTWMKLETIILSKLT 18 176 TIILSKLTQEQKSKH 18 4 LVLPSIVILDLSVEV 17 9 IVILDLSVEVLASPA 17 30 GANILRGGLSEIVLP 17 61 PPAMWTEEAGATAEA 17 67 EEAGATAEAQESGIR 17 94 GVVTEDDEAQDSIDP 17 101 EAQDSIDPPESPDRA 17 107 DPPESPDRALKAANS 17 133 VGPLWEFLLRLLKSQ 17 143 LLKSQAASGTLSLAF 17 162 LGEFLGSGTWMKLET 17 163 GEFLGSGTWMKLETI 17 172 MKLETIILSKLTQEQ 17

TABLE XLVI-V8 HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 8 SQFLEEGMGQTIPHV 24 9 QFLEEGMGGTLPHVS 24 12 EEGMGGTIPHVSPER 22 13 EGMGGTIPHVSPERV 14 7 KSQFLEEGMGGTLPH 13 2 KKGWEKSQFLEEGMG 12 6 EKSQFLEEGMGOTIP 12

TABLE XLVI-V13 HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 72 SETFLPNGINGIKDA 21 5 MGSPKSLSETFLPNG 18 1 SISMMGSPKSLSETF 16 4 MMGSPKSLSETFLPN 16 6 GSPKSLSETELPNGI 16 9 KSLSETFLPNGINGI 16 14 TELPNGINGLKDARK 16 2 ISMMGSPKSLSETFL 14 15 FLPNGINGIKDARKV 13 10 SLSETFLPNGINGIK 10

TABLE XLVI-V14 HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 10 LRLFTFWRGPVVVAA 28 2 AREIENLPLRLFTFW 25 15 FWRGPVVVAISLATF 22 13 FTFWRGPVVVAISLA 20 12 LFTFWRGPVVVAISL 18 4 EIENLPLRLFTFWRG 16 9 PLRLFTFWRGPVVVA 16 11 RLFTFWRGPVVVAIS 15 1 SAREIENLPLRLFTF 14 6 ENLPLRLFTFWRGPV 14 14 TFWRGPVVVAISLAT 14 8 LPLRLFTFWRGPVVV 12

TABLE XLVI-V21 HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 3 TIILSKLTQEQKTKH 18 2 ETIILSKLTQEQKTK 14 7 SKLTQEQKTKHCMFS 13 6 LSKLTQEQKTKHCMF 11 11 QEQKTKHCMFSLISG 11 1 LETIILSKLTQEQKT 10 9 LTQEQKTKHCMFSLI 10 10 TQEQKTKHCMFSLIS 9 12 EQKTKHCMFSLTSGS 9 5 ILSKLTQEQKTKHCM 8 8 KLTQEQKTKHCMFSL 8

TABLE XLVI-V25 HLA-DRB1-0101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 6 IILFLPGISQKLKRI 25 3 LGKIILFLPGISQKL 23 2 LLGKIILFLPGISQK 19 4 GKIILFLPGISQKLK 17 7 ILFLPCISQKLKRIK 15 9 FLPCISQKLKRIKKG 15 14 SQKLKRIKKGWEKSQ 15 15 QKLKRIKKGWEKSQF 13

TABLE XLVII-V1 HLA-DRB1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 97 HYTSLWDLRHLLVGK 28 176 QQVIELARQLNFIPI 27 228 LYSFVRDVIHPYARN 27 322 CLPMRRSERYLFLNM 27 54 YHVVIGSRNPKFASE 26 296 ETWLQCRKQLGLLSF 26 408 FHVLTYGWKRAFEEE 26 273 AGLLAAAYQLYYGTK 25 439 PSIVILDLLQLCRYP 25 109 VGKILIDVSNNMRIN 24 288 YRRFPPWLETWLQCR 24 87 NIIFVAIHREHYTSL 23 423 YYRFYTPPNFVLALV 23 133 LASLFPDSLIVKGFN 22 185 LNFIPIDLGSLSSAR 22 261 IVAITLLSLVYLAGL 22 272 LAGLLAAAYQLYYGT 22 433 VLALVLPSIVILDLL 22 145 GFNVVSAWALQLGPK 21 214 GPVVVAISLATFFFL 21 269 LVYLAGLLAAAYQLY 21 362 SFGIMSLGLLSLLAV 21 363 FGIMSLGLLSLLAVT 21 175 RQQVIELARQLNFIP 20 198 AREIENLPLRLFTLW 20 258 TLPIVAITLLSLVYL 20 264 ITLLSLVYLAGLLAA 20 376 VTSIPSVSNALNWRE 20 400 YVALLISTFHVLIYG 20 435 ALVLPSIVILDLLQL 20 438 LPSIVILDLLQLCRY 20 440 SIVILDLLQLCRYPD 20 30 KVTVGVIGSGDFAKS 19 53 GYHVVIGSRNPKFAS 19 110 GKILIDVSNNMRINQ 19 130 AEYLASLFPDSLIVK 19 151 AWALQLGPKDASRQV 19 215 PVVVAISLATFFFLY 19 217 VVAISLATFFFLYSF 19 256 NKTLPIVAITLLSLV 19 312 FAMVHVAYSLCLPMR 19 320 SLCLPMRRSERYLFL 19 402 ALLISTFHVLIYGWK 19 3 SISMMGSPKSLSETC 18 22 INGIKDARKVTVGVI 18 34 GVIGSGDFAKSLTIR 18 90 FVAIHREHYTSLWDL 18 119 NMRINQYPESNAEYL 18 139 DSLIVKGFNVVSAWA 18 143 VKGFNVVSAWALQLG 18 162 QLNFIPIDLGSLSSA 18 184 QLNFIPIDLGSLSSA 18 195 LSSAREIENLPLRLF 18 233 RDVIHPYARNQQSDF 18 308 LSFFFAMVHVAYSLC 18 331 YLFLNMAYQQVHANI 18 360 YISFGIMSLGLLSLL 18 409 HVLIYGWKRAFEEEY 18 7 MGSPKSLSETCLPNG 17 21 GINGIKDARKVTVGV 17 38 SGDFAKSLTIRLIRC 17 113 LIDVSNNMRINQYPE 17 121 RINQYPESNAEYLAS 17 155 QLGPKDASRQVYICS 17 169 SNNIQARQQVIELAR 17 178 VIELARQLNFIPIDL 17 192 LGSLSSAREIENLPL 17 225 FFFLYSFVRDVIHPY 17 249 KIPIEIVNKTLPIVA 17 292 PPWLETWLQCRKQLG 17 318 AYSLCLPMRRSERYL 17 327 RSERYLFLNMAYQQV 17 338 YQQVHANIENSWNEE 17 379 IPSVSNALNWREFSF 17 416 KRAFEEEYYRFYTPP 17 15 ETCLPNGINGIKDAR 16 72 HVVDVTHHEDALTKT 16 79 HEDALTKTNIIFVAI 16 88 IIFVAIHREHYTSLW 16 111 KILIDVSNNMRINQY 16 205 PLRLFTLWRGPVVVA 16 248 YKIPIEIVNKTLPIV 16 279 AYQLYYGTKYRRFPP 16 342 HANIENSWNEEEVWR 16 382 VSNALNWREFSFIQS 16 413 YGWKRAFEEEYYRFY 16 43 KSLTIRLIRCGYHVV 15 263 AITLLSLVYLAGLLA 15 294 WLETWLQCRKQLGLL 15 321 LCLPMRRSERYLFLN 15 367 SLGLLSLLAVTSIPS 15 387 NWREFSFIQSTLGYV 15 412 IYGWKRAFEEEYYRF 15 73 VVDVTHHEDALTKTN 14 104 LRHLLVGKILIDVSN 14 236 IHPYARNQQSDFYKI 14 267 LSLVYLAGLLAAAYQ 14 304 QLGLLSFFFAMVHVA 14 365 IMSLGLLSLLAVTSI 14 373 LLAVTSIPSVSNALN 14 401 VALLISTFHVLIYGW 14 434 LALVLPSIVILDLLQ 14 1 MESISMMGSPKSLSE 13 4 ISMMGSPKSLSETCL 13 32 TVGVIGSGDFAKSLT 13 33 VGVIGSGDFAKSLTI 13 101 LWDLRHLLVGKILID 13 138 PDSLIVKGFNVVSAW 13 164 QVYICSNNIQARQQV 13 189 PIDLGSLSSAREIEN 13 201 IENLPLRLFTLWRGP 13 213 RGPVVVAISLATFFF 13 266 LLSLVYLAGLLAAAY 13 407 TFHVLIYGWKRAFEE 13

TABLE XLVII-V2 HLA-DRB1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 6 LQALSLSLSSGFTPF 20 14 SSGFTPFSCLSLPSS 20 20 FSCLSLPSSWDYRGP 20 24 SLPSSWDYRCPPPGP 16 2 GSPGLQALSLSLSSG 12 3 SPGLQALSLSLSSGF 12 8 ALSLSLSSGFTPFSC 12 9 LSLSLSSGFTPFSCL 12 10 SLSLSSGFTPFSGLS 11 22 GLSLPSSWDYRCPPP 11 30 DYRCPPPCPADFFLY 10 31 YRCPPPCPADFFLYF 10 12 SLSSGFTPFSCLSLP 9 17 FTPFSCLSLPSSWDY 9

TABLE XLVII-V5A HLA-DRB1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 3 AREIENLPLRLFTFW 20 10 PLRLFTFWRGPVVVA 16 2 SAREIENLPLRLFTF 12 6 IENLPLRLFTFWRGP 12 8 NLPLRLFTFWRGPVV 12 5 EIENLPLRLFTFWRG 11 13 LFTFWRGPVVVAISL 10 4 REIENLPLRLFTFWR 9 11 LRLFTFWRGPVVVAI 9

TABLE XLVII-V5B HLA-DRB1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 15 IFCSFADTQTELELE 24 23 QTELELEFVFLLTLL 20 1 VSNALNWREFSFIQI 16 19 FADTQTELELEFVFL 16 21 DTQTELELEFVFLLT 16 17 CSFADTQTELELEFV 15 22 TQTELELEFVFLLTL 13 2 SNALNWREFSFIQIF 11 10 FSFIQIFCSFADTQT 11

TABLE XLVII-V6 HLA-DRB1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 8 LPSIVILGKIILFLP 26 3 VLALVLPSIVILGKI 22 9 PSIVILGKIILFLPC 22 10 SIVILGKIILFLPCI 21 17 IILFLPCISRKLKRI 20 18 ILFLPCISRKLKRIK 18 25 SRKLKRIKKGWEKSQ 18 21 LPCISRKLKRIKKGW 17 28 LKRIKKGWEKSQFLE 17 29 KRIKKGWEKSQFLEE 16 4 LALVLPSIVILGKII 14 14 LGKIILFLPCISRKL 13 15 GKIILFLPCISRKLK 13 1 NFVLALVLPSIVILG 12 5 ALVLPSIVILGKIIL 12 37 KSQFLEEGIGGTIPH 12

TABLE XLVII-V7A HLA-DRB1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 1 SISMMGSPKSLSETF 18 5 MGSPKSLSETFLPNG 17 13 ETFLPNGINGIKDAR 16 2 ISMMGSPKSLSETFL 13 12 SETFLPNG1NGIKDA 13 8 PKSLSETFLPNGING 12 4 MMGSPKSLSETFLPN 9 10 SLSETFLPNGINGIK 8

TABLE XLVII-V7B HLA-DR1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 5 YLFLNMAYQQSTLGY 18 1 RSERYLFLNMAYQQS 17 6 LFLNMAYQQSTLGYV 14 12 YQQSTLGYVALLIST 12 3 ERYLFLNMAYQQSTL 11 4 RYLFLNMAYQQSTLG 11 7 FLNMAYQQSTLGYVA 11 11 AYQQSTLGYVALLIS 11 14 QSTLGYVALLISTFH 11 8 LNMAYQQSTLGYVAL 10

TABLE XLVII-V7C HLA-DR1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 93 VGVVTEDDEAQDSID 29 130 TNGVGPLWEFLLRLL 26 7 PSIVILDLSVEVLAS 24 1 VLALVLPSIVILDLS 22 8 SIVILDLSVEVLASP 21 133 VGPLWEFLLRLLKSQ 21 3 ALVLPSIVILDLSVE 20 163 GEFLGSGTWMKLETI 20 9 IVILDLSVEVLASPA 19 123 RNPVLPHTNGVGPLW 19 137 WEFLLRLLKSQAASG 19 154 SLAFTSWSLGEFLGS 19 171 VVMKLETHLSKLTQE 19 38 LSELVLPIEWQQDRK 18 179 LSKLTQEQKSKHCMF 18 40 EIVLPIEWQQDRKIP 17 44 PIEWQQDRKIPPLST 16 90 IPVVGVVTEDDEAQD 16 176 TIILSKLTQEQKSKH 16 15 SVEVLASPAAAWKCL 15 27 KCLGANILRGGLSEI 15 32 NILRGGLSEIVLPIE 15 39 SEIVLPIEWQQDRKI 15 116 LKAANSWRNPVLPHT 15 138 EFLLRLLKSQAASGT 15 175 ETIILSKLTQEQKSK 15 2 LALVLPSIVILDLSV 14

TABLE XLVII-V8 HLA-DR1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 7 KSQFLEEGMGGTIPH 12 8 SQFLEEGMGGTIPHV 11 12 EEGMGGTLPHVSPER 10 1 IKKGWEKSQFLEEGM 9 4 GWEKSQFLEEGMGGT 7 5 WEKSQFLEEGMGGTI 7

TABLE XLVII-V13 HLA-DR1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 1 SISMMGSPKSLSETF 18 5 MGSPKSLSETFLPNG 17 13 ETFLPNGINGLKDAR 16 2 ISMMGSPKSLSETFL 13 12 SETFLPNGINGIKDA 13 8 PKSLSETFLPNGLNG 12 4 MMGSPKSLSETFLPN 9 10 SLSETFLPNGINGIK 8

TABLE XLVII-V14 HLA-DR1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 2 AREIENLPLRLFTFW 20 9 PLRLFTFWRGPVVVA 16 1 SAREIENLPLRLFTF 12 5 IENLPLRLFTFWRGP 12 7 NLPLRLETFWRGPVV 12 4 EIENLPLRLFTFWRG 11 12 LFTFWRGPVVVAISL 10 3 REIENLPLRLFTFWR 9 10 LRLETFWRGPVVVAI 9

TABLE XLVII-V21 HLA-DR1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 6 LSKLTQEQKTKHCMF 18 3 TIILSKLTQEQKTKH 16 2 ETIILSKLTQEQKTK 15 1 LETIILSKLTQEQKT 13 4 IILSKLTQEQKTKHC 10 5 ILSKLTQEQKTKHCM 9 9 LTQEQKTKHCMFSLI 9 11 QEQKTKHGMESLISG 9

TABLE XLVII-V25 HLA-DR1-0301-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 6 IILFLPCISKLKRI 21 7 ILFLPCISQKLKRIK 18 14 SQKLKRIKKGWEKSQ 18 10 LPCISQKLKRIKKGW 17 3 LGKIILFLPCISQKL 13 4 GKIILFLPCISQKLK 13 5 KIILFLPCISQKLKR 11

TABLE XLVIII-V1 HLA-DR1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 420 EEEYYRFYTPPNFVL 28 98 YTSLWDLRHLLVGKI 26 109 VGKILIDVSNNMRIN 26 175 RQQVIELARQLNFIP 26 205 PLRLFTLWRGPVVVA 26 213 RGPVVVAISLATFFF 26 225 FFFLYSFVRDVIHPY 26 229 YSFVRDVIHPYARNQ 26 312 FAMVHVAYSLCLPMR 26 370 LLSLLAVTSIPSVSN 26 373 LLAVTSIPSVSNALN 26 376 VTSIPSVSNALNWRE 26 38 SGDFAKSLTIRLIRC 22 51 RCGYHVVIGSRNPKY 22 62 NPKFASEFFPHVVDV 22 87 NIIEVAIHREHYTSL 22 143 VKGFNVVSAWALQLG 22 163 RQVYICSNNIQARQQ 22 184 QLNFIPIDLGSLSSA 22 222 LATFFFLYSFVRDVI 22 244 QSDFYKIPIEIVNKT 22 307 LLSFFFAMVHVAYSL 22 309 SFFFAMVHVAYSLCL 22 328 SERYLFLNMAYQQVH 22 346 ENSWNEEEVWRIEMY 22 357 IEMYISFGIMSLGLL 22 385 ALNWREFSFIQSTLG 22 388 WREFSFIQSTLGYVA 22 405 ISTFHVLIYGWKRAF 22 423 YYRFYTPPNFVLALV 22 429 PPNFVLALVLPSIVI 22 1 MESISMMGSPKSLSE 20 15 ETCLPNGINGIKDAR 20 19 PNGINGIKDARKVTV 20 22 INGIKDARKVTVGVI 20 30 KVTVGVIGSGDFAKS 20 47 IRLLRCGYHVVTGSR 20 53 GYHVVIGSRNPKFAS 20 70 FPHVVDVTHHEDALT 20 71 PHVVDVTHHEDALTK 20 86 TNIIFVAIHREHYTS 20 90 FVAIHREHYTSLWDL 20 101 LWDLRHLLVGKILID 20 106 HLLVGKILIDVSNNM 20 110 GKILIDVSNNMRINQ 20 111 KILIDVSNNMRINQY 20 113 LIDVSNNMRINQYPE 20 130 AEYLASLFPDSLIVK 20 133 LASLFPDSLIVKGFN 20 139 DSLIVKGFNVVSAWA 20 140 SLIVKGFNVVSAWAL 20 145 GFNVVSAWALQLGPK 20 162 SRQVYICSNNIQARQ 20 176 QQVIELARQLNFIPI 20 185 LNFIPIDLGSLSSAR 20 189 PIDLGSLSSAREIEN 20 192 LGSLSSAREIENLPL 20 217 VVAISLATFFFLYSF 20 219 AISLATFFFLYSFVR 20 233 RDVIHPYARNQQSDF 20 247 FYKIPIEIVNKTLPI 20 256 NKTLPIVAITLLSLV 20 258 TLPIVAITLLSLVYL 20 261 IVAITLLSLVYLAGL 20 264 ITLLSLVYLAGLLAA 20 266 LLSLVYLAGLLAAAY 20 267 LSLVYLAGLLAAAYQ 20 273 AGLLAAAYQLYYGTK 20 292 PPWLETWLQCRKQLG 20 302 RKQLGLLSFFFAMVH 20 304 QLGLLSFFFAMVHVA 20 331 YLFLNMAYQQFHANI 20 351 EEEVWRIEMYISFGI 20 354 VWRIEMYISFGIMSL 20 362 SFGIMSLGLLSLLAV 20 365 IMSLGLLSLLAVTSI 20 367 SLGLLSLLAVTSIPS 20 368 LGLLSLLAVTSIPSV 20 379 IPSVSNALNWREFSF 20 395 QSTLGYVALLISTFH 20 398 LGYVALLISTFHVLI 20 401 VALLISTFHVLIYGW 20 430 PNFVLALVLPSIVIL 20 431 NFVLALVLPSIVILD 20 435 ALFLPSIVILDLLQL 20 438 LPSIVILDLLQLCRY 20 440 SIVILDLLQLCRYPD 20 12 SLSETCLPNGINGIK 18 21 GINGIKDARDVTVGV 18 36 IGSGDFAKSLTIRLI 18 76 VTHHEDALTKTNIIF 18 97 HYTSLWDLRHLLVGK 18 142 IVKGFNVVSAWALQL 18 154 LQLGPKDASRQVYIC 18 161 ASRQVYICSNNIQAR 18 168 CSNNIQARQQVIELA 18 186 NFLPIDLGSLSSARE 18 195 LSSARELENLPLRLF 18 234 DVIHPYARNQQSDFY 18 248 YKIPIEIVNKTLPIV 18 257 KTLPIVAITLLSLVY 18 289 RRFPPWLETWLQCRK 18 339 QQVHANIENSWNEEE 18 348 SWNEEEVWRIEMYIS 18 359 MYISFGIMSLGLLSL 18 364 GIMSLGLLSLLAVTS 18 384 NALNWREFSFIQSTL 18 387 NWREFSFIQSTLGYV 18 399 GYVALLISTFHVLIY 18 432 FVLALVLPSIVILDL 18 66 ASEFFPHVVDVTHHE 16 67 SEFFPHVVDVTHHED 16 95 REHYTSLWDLRHLLV 16 122 INQYPESNAEYLASL 16 129 NAEYLASLFPDSLIV 16 206 LRLFTLWRGPVVVAI 16 209 FTLWRGPVVVAISLA 16 224 TFFFLYSFVRDVIHP 16 226 FFLYSFVRDVIHPYA 16 228 LYSFVRDVIHPYARN 16 236 IHPYARNQQSDFYKI 16 245 SDFYKIPIEIVNKTL 16 268 SLVYLAGLLAAAYQL 16 285 GTKYRRFPPWLETWL 16 288 YRRFPPWLETWLQCR 16 308 LSFFFAMVHVAYSLC 16 330 RYLFLNMAYQQVHAN 16 335 NMAYQQVHANIENSW 16 352 EEVWRIEMYISFGIM 16 360 YISFGIMSLGLLSLL 16 390 EFSFIQSTLGYVALL 16 397 TLGYVALLISTFHVL 16 412 IYGWKRAFEEEEYRF 16 416 KRAFEEEYYRFYTPP 16 424 YRFYTPPNFVLALVL 16 296 ETWLQCRKQLGLLSF 15 3 SISMMGSPKSLSETC 14 4 ISMMGSPKSLSETCL 14 32 TVGVIGSGDFAKSLT 14 33 VGVIGSGDFAKSLTI 14 44 SLTIRLIRCGYHVVI 14 46 TIRLIRCGYHVVIGS 14 54 YHVVIGSRNPKFASE 14 73 VVDVTHHEDALTKTN 14 80 EDALTKTNIIFVAIH 14 85 KTNIIFVAIHREHYT 14 88 IIFVAIHREHYTSLW 14 117 SNNMRINQYPESNAE 14 119 NMRINQYPESNAEYL 14 151 AWALQLGPKDASRQV 14 178 VIELARQLNFIPIDL 14 182 ARQLNFIPIDLGSLS 14 187 FIPIDLGSLSSAREI 14 198 AREIENLPLRLFTLW 14 203 NLPLRLFTLWRGPVV 14 208 LFTLWRGPVVVAISL 14 214 GPVVVAISLATFFFL 14 232 VRDVIHPYARNQQSD 14 249 KIPIEIVNKTLPIVA 14 252 IEIVNKTLPIVAITL 14 259 LPIVAITLLSLVYLA 14 263 AITLLSLVYLAGLLA 14 269 LVYLAGLLAAAYQLY 14 272 LAGLLAAAYQLYYGT 14 305 LGLLSFFFAMVHVAY 14 311 FFAMVHVAYSLCLPM 14 314 MVHVAYSLCLPMRRS 14 318 AYSLCLPMRRSERYL 14 322 CLPMRRSERYLFLNM 14 329 ERYLFLNMAYQQVHA 14 333 FLNMAYQQVHAINEN 14 342 HANIENSWNEEEVWR 14 356 RIEMYISFGIMSLGL 14 363 FGIMSLGLLSLLAVT 14 371 LSLLAVTSIPSVSNA 14 391 FSFIQSTLGYVALLI 14 400 VYALLISTFHVLIYG 14 402 ALLISTFHVLIYGWK 14 407 TFHVLIYGWKRAFEE 14 409 HVLIYGWKRAFEEEY 14 433 VLALVLPSIVILDLL 14 439 PSIVILDLLQLCRYP 14

TABLE XLVIII-V5A HLA-DR1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 14 SSGETPFSCLSLPSS 22 17 FTPFSCLSLPSSWDY 22 3 SPGLQALSLSLSSGF 20 10 SLSLSSGFTPFSCLS 20 2 GSPGLQALSLSLSSG 18 7 QALSLSLSSGFTPFS 18 28 SWDYRCPPPCPADFF 18 6 LQALSLSLSSGFTPF 14 20 FSCLSLPSSWDYRCP 14 4 PGLQALSLSLSSGFT 12 13 LSSGFTPFSCLSLPS 12 16 GFTPFSCLSLPSSWD 12 19 PFSCLSLPSSWDYRC 12 24 SLPSSWDYRCPPPCP 12

TABLE XLVIII-V5B HLA-DR1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 4 ALNWREFSFIQIFGS 22 7 WREFSFIQIFCSFAD 22 9 EFSFIQLFCSFADTQ 22 13 IQIFGSFADTQTELE 22 10 FSFIQIFCSFADTQT 20 23 QTELELEFVFLLTLL 20 3 NALNWREFSFIQIFC 18 15 IFCSFADTQTELELE 18 16 FCSFADTQTELELEF 16 12 FIQIFGSFADTQTEL 14 6 NWREFSFIQIFCSFA 12 14 QIFCSFADTQTELEL 12 20 ADTQTELELEFVFLL 12 22 TQTELELEFVFLLTL 12 24 TELELEFVFLLTLLL 12

TABLE XLVIII-V6 HLA-DR1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 18 ILFLPCISRKLKRIK 26 17 IILFLPGISRKLKRT 22 37 KSQFLEEGIGGTIPH 22 1 NFVLALVLPSIVILG 20 5 ALVLPSIVILGKIIL 20 8 LPSIVILGKIILFLP 20 14 LGKIILFLPCISRKL 20 46 GGTIPHVSPERVTVM 20 2 FVLALVLPSIVILGK 18 22 PCISRKLKRIKKGWE 18 30 RIKKGWEKSQFLEEG 18 3 VLALVLPSIVILGKI 14 11 IVILGKIILFLPGIS 14 15 GKIILFLPCISRKLK 14 16 KIILFLPGISRKLKR 14 25 SRKLKRIKKGWEKSQ 14 28 LKRIKKGWEKSQFLE 14 38 SQFLEEGIGGTLPHV 14 42 EEGIGGTIPHVSPER 14 6 LVLPSIVILGKIILF 12 7 VLPSIVILGKIILFL 12 13 ILGKIILFLPGISRK 12 34 GWEKSQFLEEGIGGT 12 43 EGIGGTIPHVSPERV 12

TABLE XLVIII-V7A HLA-DR1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 13 ETFLPNGINGIKDAR 20 10 SLSETFLPNGINGIK 18 12 SETFLPNGINGIKDA 16 1 SISMMGSPKSLSETF 14 2 ISMMGSPKSLSETFL 14 5 MGSPKSLSETFLPNG 12 7 SPKSLSETFLPNGIN 12 9 KSLSETFLPNGINGI 12

TABLE XLVIII-V7B HLA-DR1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 5 YLFLNMAYQQSTLGY 26 2 SERYLFLNMAYQQST 22 14 QSTLGYVALLISTFH 20 4 RYLFLNMAYQQSTLG 16 9 NMAYQQSTLGYVALL 16 3 ERYLFLNMAYQQSTL 14 7 FLNMAYQQSTLGYVA 14 1 RSERYLFLNMAYQQS 12 6 LFLNMAYQQSTLGYV 12 11 AYQQSTLGYVALLIS 12 15 STLGYVALLISTFHV 12

TABLE XLVIII-V7C HLA-DR1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 134 GPLWEFLLRLLKSQA 28 168 SGTWMKLETIILSKL 28 7 PSIVILDLSVEVLAS 26 13 DLSVEVLASPAAAWK 26 113 DRALKAANSWRNPVL 26 138 EFLLRLLKSQAASGT 26 150 SGTLSLAFTSWSLGE 26 176 TIILSKLTQEQKSKH 26 23 AAAWKCLGANILRGG 22 62 PAMWTEEAGATAEAQ 22 162 LGEFLGSGTWMKLET 22 3 ALVLPSIVILDLSVE 20 8 SIVILDLSVEVLASP 20 31 ANILRGGLSEIVLPI 20 40 EIVLPIEWQQDRKVV 20 50 DRKIPPLSTPPPPAM 20 61 PPAMWTEEAGATAEA 20 89 QIPVVGVVTEDDEAQ 20 92 VVGVVTEDDEAQDSI 20 130 TNGVGPLWEFLLRLL 20 133 VGPLWEFLLRLLKSQ 20 137 WEFLLRLLKSQAASG 20 159 SWSLGEFLGSGTWMK 20 169 GTWMKLETIILSKLT 20 171 WMKLETIILSKLTQE 20 27 KCLGANILRGGLSEI 18 74 EAQESGIRNKSSSSS 18 95 VVTEDDEAQDSIDPP 18 142 RLLKSQAASGTLSLA 18 151 GTLSLAFTSWSLGEF 18 172 MKLETIILSKLTQEQ 18 44 PIEWQQDRKIPPLST 16 119 ANSWRNPVLPHTNGV 16 157 FTSWSLGEFLGSGTW 16 77 ESGLRNKSSSSSQIP 15 175 ETIILSKLTQEQKSK 15 1 VLALVLPSIVILDLS 14 6 LPSIVILDLSVEVLA 14 9 IVLLDLSVEVLASPA 14 11 ILDLSVEVLASPAAA 14 16 VEVLASPAAAWKGLG 14 30 GANILRGGLSEIVLP 14 35 RGGLSEIVLPIEWQQ 14 38 LSEIVLPIEWQQDRK 14 39 SEIVLPIEWQQDRKI 14 42 VLPIEWQQDRKIPPL 14 53 IPPLSTPPPPAMWTE 14 87 SSQIPVVGVVTEDDE 14 90 IPVVGVVTEDDEAQD 14 93 VGVVTEDDEAQDSID 14 103 QDSIDPPESPDRALK 14 123 RNPVLPHTNGVGPLW 14 141 LRLLKSQAASGTLSL 14 163 GEFLGSGTWMKLETI 14 179 LSKLTQEQKSKHCMF 14

TABLE XLVIII-V8 HLA-DR1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 7 KSQFLEEGMGGTIPH 22 8 SQFLEEGMGGTIPHV 14 12 EEGMGGTIPHVSPER 14 4 GWEKSQFLEEGMGGT 12 13 EGMGGTIPHVSPERV 12 2 KKGWEKSQFLEEGMG 10

TABLE XLVIII-V13 HLA-DRB1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 13 ETFLPNGINGIKDAR 20 10 SLSETFLPNGINGIK 18 12 SETFLPNGJNGIKDA 16 1 SISMMGSPKSLSETF 14 2 ISMMGSPKSLSETFL 14 5 MGSPKSLSETFLPNG 12 7 SPKSLSETFLPNGIN 12 9 KSLSETFLPNGINGI 12

TABLE XLVIII-V14 HLA-DRB1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 9 PLRLFTFWRGPVVVA 26 10 LRLFTFWRGPVVVAI 16 12 LFTFWRGPVVVAISL 16 13 FTFWRGPVVVAISLA 16 2 AREIENLPLRLFTFW 14 7 NLPLRLFTFWRGPVV 14 3 REIENLPLRLFTFWR 12 6 ENLPLRLFTFWRGPV 12 14 TFWRGPVVVAISLAT 12 15 FWRGPVVVAISLATF 12

TABLE XLVIII-V21 HLA-DRB1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 3 TIILSKLTQEQKTKH 26 2 ETIILSKLTQEQKTK 15 6 LSKLTQEQKTKHGMF 14 5 ILSKLTQEQKTKHCM 12

TABLE XLVIII-V25 HLA-DRB1-0401-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 7 ILFLPCISQKLKRIK 26 6 IILFLPGISQKLKRI 22 3 LGKIILFLPCISQKL 20 4 GKIILFLPCISQKLK 20 11 PCISQKLKRIKKGWE 18 5 KIILFLPCISQKLKR 14 14 SQKLKRLKKGWEKSQ 14 2 ILGKIILFLPCISQK 12

TABLE XLIX-V1 HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 249 KIPIEIVNKTLPPVA 27 308 LSFFFAMVHVAYSLC 27 229 YSFVRDVIHPYARNQ 26 281 QLYYGTKYRRFPPWL 25 295 LETWLQGRKQLGLLS 25 87 NIIFVAIHREHYTSL 24 388 WREFSFIQSTLGYVA 23 309 SFFFAMVHVAYSLCL 22 3 SISMMGSPKSLSETG 21 71 PHVVDVTHHEDALTK 21 98 YTSLWDLRHLLVGKI 21 175 RQQVIELARQLNFIP 21 205 PLRLFTLWRGPVVVA 21 70 FPHVVDVTHHEDALT 20 95 REHYTSLWDLRHLLV 20 151 AWALQLGPKDASRQV 20 263 AITLLSLVYLAGLLA 20 1 MESISMMGSPKSLSE 19 51 RCGYHVVIGSRNPKF 19 106 HLLVGKILIDVSNNM 19 182 ARQLNFIPIDLGSLS 19 266 LLSLVYLAGLLAAAY 19 351 EEEVWRIEMYISFGI 19 395 QSTLGYVALLISTFH 19 424 YRFYTPPNFVLALVL 19 67 SEFFPHVVDVTHHED 18 222 LATFFFLYSFVRDVI 18 302 RKQLGLLSFFFAMVH 18 307 LLSFFFAMVHVAYSL 18 367 SLGLLSLLAVTSIPS 18 370 LLSLLAVTSIPSVSN 18 28 ARKVTVGVIGSGDFA 17 86 TNIIFVAIHREHYTS 17 99 TSLWDLRHLLVGKIL 17 134 ASLFPDSLIVKGFNV 17 143 VKGFNVVSAWALQLG 17 225 FFFLYSFVRDVIHPY 17 226 FFLYSFVRDVIHPYA 17 244 QSDFYKIPIEIVNKT 17 335 NMAYQQVHANIENSW 17 360 YISFGIMSLGLLSLL 17 405 ISTFHVLIYGWKRAF 17 129 NAEYLASLFPDSLIV 16 136 LFPDSLIVKGFNVVS 16 163 RQVYICSNNIQARQQ 16 184 QLNFIPIDLGSLSSA 16 268 SLVYLAGLLAAAYQL 16 279 AYQLYYGTKYRRFPP 16 282 LYYGTKYRRFPPWLE 16 328 SERYLFLNMAYQQVH 16 330 RYLFLNMAYQQVHAN 16 385 ALNWREFSFIQSTLG 16 397 TLGYVALLISTFHVL 16 429 PPNFVLALVLPSIVI 16 42 AKSLTIRLIRCGYHV 15 47 IRLIRCGYHVVIGSR 15 103 DLRHLLVGKILIDVS 15 142 IVKGFNVVSAWALQL 15 210 TLWRGPVVVAISLAT 15 317 VAYSLCLPMRRSERY 15 318 AYSLCLPMRRSERYL 15 322 CLPMRRSERYLFLNM 15 401 VALLISTFHVLIYGW 15 408 FHVLIYGWKRAFEEE 15 428 TPPNFVLALVLPSIV 15 19 PNGINGIKDARKVTV 14 22 INGIKDARKVTVGVI 14 43 KSLTIRLIRCGYHVV 14 52 CGYHVVIGSRNPKFA 14 53 GYHVVIGSRNPKFAS 14 56 VVIGSRNPKFASEFF 14 66 ASEFFPHVVDVTHHE 14 77 THHEDALTKTNIIFV 14 85 KTNIIFVAIHREHYT 14 89 IFVAIHREHYTSLWD 14 113 LIDVSNNMRINQYPE 14 189 PIDLGSLSSAREIEN 14 198 AREIENLPLRLFTLW 14 203 NLPLRLFTLWRGPVV 14 212 WRGPVVVAISLATFF 14 233 RDVIHPYARNQQSDF 14 261 IVAITLLSLVYLAGL 14 319 YSLCLPMRRSERYLF 14 348 SWNEEEVWRIEMYIS 14 373 LLAVTSIPSVSNALN 14 381 SVSNALNWREFSFIQ 14 407 TFHVLIYGWKRAFEE 14 409 HVLIYGWKRAFEEEY 14 430 PNFVLALVLPSIVIL 14 435 ALVLPSIVILDLLQL 14 30 KVTVGVIGSGDFAKS 13 33 VGVIGSGDFAKSLTI 13 101 LWDLRHLLVGKILID 13 139 DLSIVKGFNVVSAWA 13 146 FNVVSAWALQLGPKD 13 178 VIELARQLNFIPIDL 13 185 LNFIPIDLGSLSSAR 13 206 LRLFTLWRGPVVVAI 13 208 LFTLWRGPVVVAISL 13 223 ATFFFLYSFVRDVIH 13 252 IEIVNKTLPIVAITL 13 256 NKTLPIVAITLLSLV 13 280 YQLYYGTKYRRFPPW 13 311 FFAMVHVAYSLCLPM 13 358 EMYISFGIMSLGLLS 13 364 GIMSLGLLSLLAVTS 13 376 VTSIPSVSNALNWRE 13 391 FSFIQSTLGYVALLI 13 431 NFVLALVLPSIVILD 13

TABLE XLIX-V2 HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 17 FTPFSCLSLPSSWDY 22 3 SPGLQALSLSLSSGF 19 28 SWDYRCPPPCPADFF 16 24 SLPSSWDYRCPPPCP 14 5 GLQALSLSLSSGFTP 12 8 ALSLSLSSGFTPFSG 12 10 SLSLSSGFTPFSCLS 12 14 SSGFTPFSGLSLPSS 12 26 PSSWDYRCPPPCPAD 10

TABLE XLIX-V5A HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 13 LFTFWRGPVVVAISL 17 10 PLRLETFWRGPVVVA 15 15 TFWRGPVVVAISLAT 15 3 AREIENLPLRLFTFW 14 8 NLPLRLFTFWRGPVV 14 11 LRLFTFWRGPVVVAI 13 14 FTFWRGPVVVAISLA 12 16 FWRGPVVVAISLATF 9 4 REIENLPLRLFTFWR 8

TABLE XLIX-V5B HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 7 WREFSFIQIFGSFAD 22 9 EFSFIQIFCSFADTQ 22 16 FCSFADTQTELELEF 11 4 ALNWREFSFIQIFCS 10 13 IQIECSFADTQTELE 10

TABLE XLIX-V6 HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 8 LPSIVILGKIILFLP 21 18 ILFLPGISRKLKRIK 21 25 SRKLKRIKKGWEKSQ 20 43 EGIGGTIPHVSPERV 20 11 IVILGKIILFLPCIS 19 21 LPCISRKLKRIKKGW 16 22 PCISRKLKRIKKGWE 15 5 ALVLPSIVILGKIIL 14 46 GGTIPHVSPERVTVM 14 1 NFVLALVLPSIVILG 13 4 LALVLPSIVILGKII 13 14 LGKIILFLPCISRKL 13 35 WEKSQFLEEGIGGTI 13 39 QFLEEGIGGTIPHVS 13 42 EEGIGGTIPHVSPER 13 15 GKIILFLPCISRKLK 12 17 IILFLPCISRKLKRI 12 32 KKGWEKSQFLEEGIG 10 37 KSQFLEEGIGGTIPH 10

TABLE XLIX-V7A HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 1 SISMMGSPKSLSETF 21 8 PKSLSETFLPNGING 12 12 SETFLPNGINGIKDA 10

TABLE XLIX-V7B HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 4 RYLFLNMAYQQSTLG 22 14 QSTLGYVALLISTFH 19 2 SERYLFLNMAYQQST 16 7 FLNMAYQQSTLGYVA 13 9 NMAYQQSTLGYVALL 10

TABLE XLIX-V7C HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 137 WEFLLRLLKSQAASG 26 134 GPLWEFLLRLLKSQA 25 44 PIEWQQDRKJPPLST 24 121 SWRNPVLPHTNGVGP 21 13 DLSVEVLASPAAAWK 19 50 DRKIPPLSTPPPPAM 18 62 PAMWTEEAGATAEAQ 18 138 EFLLRLLKSQAASGT 18 23 AAAWKCLGANILRGG 17 168 SGTWMKLETIILSKL 17 179 LSKLTQEQKSKHCMF 17 157 FTSWSLGEFLGSGTW 16 9 IVILDLSVEVLASPA 15 11 ILDLSVEVLASPAAA 15 19 LASPAAAWKCLGANI 15 35 RGGLSEIVLPIEWQQ 15 43 LPIEWQQDRKIPPLS 15 73 AEAQESGIRNKSSSS 15 3 ALVLPSIVILDLSVE 14 27 KCLGANILRGGLSEI 14 75 AQESGIRNKSSSSSQ 14 89 QIPVVGVVTEDDEAQ 14 135 PLWEFLLRLLKSQAA 14 173 KLETIILSKLTQEQK 14 4 LVLPSIVILDLSVEV 13 6 LPSIVILDLSVEVLA 13 8 SIVILDLSVEVLASP 13 26 WKCLGANILRGGLSE 13 28 CLGANILRGGLSEIV 13 87 SSQIPVVGVVTEDDE 13 90 IPVVGVVTEDDEAQD 13 123 RNPVLPHTNGVGPLW 13 130 TNGVGPLWEFLLRLL 13 152 TLSLAFTSWSLGEFL 13 156 AFTSWSLGEFLGSGT 13 169 GTWMKLETIILSKLT 13 171 WMKLETIILSKLTQE 13 10 VILDLSVEVLASPAA 12 12 LDLSVEVLASPAAAW 12 39 SEIVLPIEWQQDRKI 12 58 TPPPPAMWTEEAGAT 12 74 EAQESGIRNKSSSSS 12 77 ESGIRNKSSSSSQIP 12 100 DEAQDSIDPPESPDR 12 110 ESPDRALKAANSWRN 12 119 ANSWRNPVLPHTNGV 12 124 NPVLPHTNGVGPLWE 12 140 LLRLLKSQAASGTLS 12 150 SGTLSLAFTSWSLGE 12 154 SLAFTSWSLGEFLGS 12 176 TIILSKLTQEQKSKH 12

TABLE XLIX-V8 HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 13 EGMGGTIPHVSPERV 20 9 QFLEEGMGGTIPHVS 13 12 EEGMGGTIPHVSPER 13 5 WEKSQFLEEGMGGTI 12 2 KKGWEKSQFLEEGMG 10 7 KSQFLEEGMGGTIPH 10

TABLE XLIX-V13 HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 27; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 1 SISMMGSPKSLSETF 21 8 PKSLSETFLPNGING 12 12 SETFLPNGINGIKDA 10

TABLE XLIX-V14 HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 29; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 12 LFTFWRGPVVVAISL 17 9 PLRLFTFWRGPVVVA 15 14 TFWRGPVVVAISLAT 15 2 AREIENLPLRLFTFW 14 7 NLPLRLFTFWRGPVV 14 10 LRLFTFWRGPVVVAI 13 13 FTFWRGPVVVAISLA 12 15 FWRGPVVVAISLATF 9 3 REIENLPLRLFTFWR 8

TABLE XLIX-V21 HLA-DRB1-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 6 LSKLTQEQKTKHGMF 17 3 TIILSKLTQEQKTKH 12 8 KLTQEQKTKHCMFSL 8

TABLE XLIX-V21 HLA-DRB-1101-15mers-98P4B6 Each peptide is a portion of SEQ ID NO: 43; each start position is specified, the length of peptide is 15 amino adds, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 9 LTEQKTKHGMFSLI 8

TABLE XLIX-V25 HLA-DRB1-1101-15mers-98P4B6 Each peplide is a portion of SEQ ID NO: 51; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen. Pos 123456789012345 score 14 SQKLKRLKKGWEKSQ 20 10 LPCISQKLKRIKKGW 16 11 PCISQKIKRIXKGWE 15 3 LGKIILFLPCISQKL 13 7 ILFLPCISQKLKRIK 13 4 GKIHLFLPCISQKLK 12 6 IILFLPCISQKLKRI 11 8 LFLPCISQKLKRKK 9

TABLE L Properties of 98P4B6 Bioinformatic Program Outcome V.1 ORF ORF finder Protein length 454 aa Transmembrane region TM Pred 6TM, aa 214-232, 261-286, 304-325, 359-379, 393-415, 426-447, N-term inside HMMTop 6TM, aa 215-232 261-279 306-325 360-379 396-415 428-447 N-term ou Sosui 6TM, aa 206-228, 255-277, 304-325, 359-381, 393-415, 428-450 TMHMM 6TM, aa 210-232, 262-284, 304-323, 360-382, 392-414, 427-449 Signal Peptide Signal P none pI pI/MW tool pI 8.74 Molecular weight pI/MW tool 52.0 kD Localization PSORT Plasma membrane 60%, golgi 40% PSORT II Endoplasmic reticulum 39%, plasma membrane 34% Motifs Pfam no known motifs Prints pyridine nucleotide reductase ProDom Dudulin, oxidoreductase Blocks adenosyl-L-homocysteine hydrolase V.2 ORF ORF finder Protein length 45 aa Transmembrane region TM Pred 1 TM, aa 5-23, N-term inside HMMTop no TM Sosui souble protein TMHMM no TM Signal Peptide Signal P none pI pI/MW tool pI 4.2 Molecular weight pI/MW tool 4.84 kD Localization PSORT Ouside 37%, microbody 32% PSORT II Extracellular 33%, nuclear 33% Motifs Pfam no known motifs Prints no known motifs Blocks no known motifs V.5 ORF ORF finder Protein length 419 aa Transmembrane region TM Pred 4TM, aa 214-232, 261-286, 304-325, 359-379 N-term inside HMMTop 4TM, aa 215-232, 259-278, 305-324, 360-379 N-term outside Sosui 4TM, aa 209-231, 255-277, 304-325, 356-379 TMHMM 4TM, aa 210-232, 262-284, 304-323, 360-382 Signal Peptide Signal P none pI pI/MW tool pI 8.1 Molecular weight pI/MW tool 47.9 kD Localization PSORT Plasma membrane 60%, golgi 40% PSORT II Endoplasmic reticulum 44%, plasma membrane 22% Motifs Pfam no known motifs Prints no known motifs ProDom Dudulin, oxidoreductase Blocks no known motifs V.6 ORF ORF finder Protein length 490 aa Transmembrane region TM Pred 6TM, aa 214-232, 261-286, 304-325, 359-379, 393-415, 432-455 HMMTop 7TM, aa 140-158, 214-232, 259-280, 305-323, 361-383, 396-413, 432-455, N-term out Sosui 6TM, aa 206-228, 255-277, 304-325, 359-381, 393-415, 428-450 TMHMM 6TM, aa 210-232, 262-284, 304-323, 360-382, 392-414, 427-449 Signal Peptide Signal P none pI pI/MW tool pI 9.2 Molecular weight pI/MW tool 55.9 kD Localization PSORT Plasma membrane 60%, golgi 40% PSORT II Endoplasmic reticulum 39%, plasma membrane 34% Motifs Pfam no known motifs Prints pyridine nucleotide reductase ProDom Dudulin, oxidoreductase Blocks adenosyl-L-homocysteine hydrolase V.7 ORF ORF finder Protein length 576 aa Transmembrane region TM Pred 6TM, aa 214-232, 262-280, 306-322, 331-360, 371-393, 525-544. N-term out HMMTop 5TM, aa 215-232, 261-279, 306-325, 342-359, 378-397 N- term out Sosui 5 TM, aa 206-228, 255-277, 304-325, 339-360, 380-402 TMHMM 4TM, aa 210-232, 262-284, 304-323, 343-360 Signal Peptide Signal P none pI pI/MW tool pI 8.5 Molecular weight pI/MW tool 64.5 kD Localization PSORT Plasma membrane 60%, golgi 40% PSORT II Endoplasmic reticulum 44%, plasma membrane 22% Motifs Pfam no known motifs Prints pyridine nucleotide reductase ProDom Dudulin, oxidoreductase Blocks Ets domain, adenosyl-L-homocysteine hydrolase

TABLE LI Exon boundaries of transcript 98P4B6 v.1 Exon Number Start End Length 1 23 321 299 2 322 846 525 3 847 1374 528 4 1375 1539 165 5 1540 1687 148 6 1688 2453 766

TABLE LII(a) Nucleotide sequence (partial, 5′ open) of transcript variant 98P4B6 v.2 (SEQ ID NO: 153) agtggatccc ccgggctgca ggctctctct ctctctctct   60 cttccgggtt cacgccattc tcctgcctca gcctcccgag tagctgggac tacaggtgcc  120 cgccaccatg cccggctgat ttctttttgt atttttagta cagacggagt ttcaccgtgt  180 tagccaggat ggtctcgatc tcctgacctc gtgatccgcc cgccttggcc tccaaagtgc  240 tgggattaca ggtgtgagct accgcgcccg gcctattatc ttgtactttc taactgagcc  300 ctctattttc tttattttaa taatatttct ccccacttga gaatcacttg ttagttcttg  360 gtaggaattc agttgggcaa tgataacttt tatgggcaaa aacattctat tatagtgaac  420 aaatgaaaat aacagcgtat tttcaatatt ttcttattcc tttaaattcca ctctttaac  480 actatgctta accacttaat gtgatgaaat attcctaaaa gttaaatgac tattaaagca  540 tatattgttg catgtatata ttaagtagcc gatactctaa ataaaaatac cactgttaca  600 gataaatggg gcctttaaaa atatgaaaaa caaacttgtg aaaatgtata aaagatgcat  660 ctgttgtttc aaatggcact atcttctttt cagtactaca aaaacagaat aattttgaag  720 ttttagaata aatgtaatat atttactata attctaaatg tttaaatgct tttctaaaaa  780 tgcaaaacta tgatgtttag ttgctttatt ttacctctat gtgattattt ttcttaattg  840 ttatttttta taatcattat ttttctgaac cattcttctg gcctcagaag taggactgaa  900 ttctactatt gctaggtgtg agaaagtggt ggtgagaacc ttagagcagt ggagatttgc  960 tacctggtct gtgttttgag aagtgcccct tagaaagtta aaagaatgta gaaaagatac 1020 tcagtcttaa tcctatgcaa aaaaaaaatc aagtaattgt tttcctatga ggaaaataac 1080 catgagctgt atcatgctac ttagctttta tgtaaatatt tcttatgtct cctctattaa 1140 gagtatttaa aatcatattt aaatatgaat ctattcatgc taacattatt tttcaaaaca 1200 tacatggaaa tttagcccag attgtctaca tataaggttt ttatttgaat tgtaaaatat 1260 ttaaaagtat gaataaaata tatttatagg tatttatcag agatgattat tttgtgctac 1320 atacaggttg gctaatgagc tctagtgtta aactacctga ttaatttctt ataaagcagc 1380 ataaccttgg cttgattaag gaattctact ttcaaaaatt aatctgataa tagtaacaag 1440 gtatattata ctttcattac aatcaaatta tagaaattac ttgtgtaaaa gggcttcaag 1500 aatatatcca atttttaaat attttaatat atctcctatc tgataactta attcttctaa 1560 attaccactt gccattaagc tatttcataa taaattctgt acagtttccc ccaaaaaaag 1620 agatttattt atgaaatatt taaagtttct aatgtggtat tttaaataaa gtatcataaa 1680 tgtaataagt aaatatttat ttaggaatac tgtgaacact gaactaatta ttcctgtgtc 1740 agtctatgaa atccctgttt tgaaataagt aaacagccta aaatgtgttg aaattatttt 1800 gtaaatccat gacttaaaac aagatacata catagtataa cacacctcac agtgttaaga 1860 tttatattgt gaaatgagac accctacctt caattgttca tcagtgggta aaacaaattc 1920 tgatgtacat tcaggacaaa tgattagccc taaatgaaac tgtaataatt tcagtggaaa 1980 ctcaatctgt ttttaccttt aaacagtgaa ttttacatga atgaatgggt tcttcacttt 2040 ttttttagta tgagaaaatt atacagtgct taattttcag agattctttc catatgttac 2100 taaaaaatgt tttgttcagc ctaacatact gagttttttt taactttcta aattattgaa 2160 tttccatcat gcattcatcc aaaattaagg cagactgttt ggattcttcc agtggccaga 2220 tgagctaaat taaatcacaa aagcagatgc ttttgtatga tctccaaatt gccaacttta 2280 aggaaatatt ctcttgaaat tgtctttaaa gatcttttgc agctttgcag atacccagac 2340 tgagctggaa ctggaatttg tcttcctatt gactctactt ctttaaaagc ggctgcccat 2400 tacattcctc agctgtcctt gcagttaggt gtacatgtga ctgagtgttg gccagtgaga 2460 tgaagtctcc tcaaaggaag gcagcatgtg tcctttttca tcccttcatc ttgctgctgg 2520 gattgtggat ataacaggag ccctggcagc tgtctccaga ggatcaaagc cacacccaaa 2580 gagtaaggca gattagagac cagaaagacc ttgactactt ccctacttcc actgcttttt 2640 cctgcattta agccattgta aatctgggtg tgttacatga agtgaaaatt aattctttct 2700 gcccttcagt tctttatcct gataccattt aacactgtct gaattaacta gactgcaata 2760 attctttctt ttgaaagctt ttaaaggata atgtgcaatt cacattaaaa ttgattttcc 2820 attgtcaatt agttatactc attttcctgc cttgatcttt cattagatat tttgtatctg 2880 cttggaatat attatcttct ttttaactgt gtaattggta attactaaaa ctctgtaatc 2940 tccaaaatat tgctatcaaa ttacacacca tgttttctat cattctcata gatctgcctt 3000 ataaacattt aaataaaaag tactatttaa tgatttaaaa aaaaaaaaaa aaaaaaaaaa a 3041

TABLE LIII(a) Nucleotide sequence alignment of 98P4B6 v.1 (SEQ ID NO: 154) and 98P4B6 v.2 (SEQ ID NO: 155) Score = 1429 bits (743), Expect = 0.0 Identities = 750/751 (99%), Gaps = 1/751 (0%) Strand = Plus/Plus

NOTE: THERE WAS A SINGLE NUCLEOTIDE INSERTION OF A SINGLE BASE AT 2620 OF V.2.

TABLE LIV(a) Peptide sequences (partial) of protein coded by 98P4B6 v.2 (SEQ ID NO: 156) SGSPGLQALS LSLSSGFTPF SCLSLPSSWD YRCPPPCPAD 45 FFLYF

TABLE LV(a) Amino acid sequence alignment of 98P4B6 v.1 and 98P4B6 v.2 --NO SIGNIFICANT HOMOLOGY--

TABLE LII(b) Nucleotide sequence of transcript variant 98P4B6 v.3 (SEQ ID NO: 157) ttctgctata gagatggaac agtatatgga aagctcccaa   60 gaaagtgaag agaggaaatt ggaaaattgt gagtggacct tctgatactg ctcctccttg  120 cgtggaaaag gggaaagaac tgcatgcata ttattcagcg tcctatattc aaaggatatt  180 cttggtgatc ttggaagtgt ccgtatcatg gaatcaatct ctatgatggg aagccctaag  240 agccttagtg aaacttgttt acctaatggc ataaatggta tcaaagatgc aaggaaggtc  300 actgtaggtg tgattggaag tggagatttt gccaaatcct tgaccattcg acttattaga  360 tgcggctatc atgtggtcat aggaagtaga aatcctaagt ttgcttctga attttttcct  420 catgtggtag atgtcactca tcatgaagat gctctcacaa aaacaaatat aatatttgtt  480 gctatacaca gagaacatta tacctccctg tgggacctga gacatctgct tgtgggtaaa  540 atcctgattg atgtgagcaa taacatgagg ataaaccagt acccagaatc caatgctgaa  600 tatttggctt cattattccc agattctttg attgtcaaag gatttaatgt tgtctcagct  660 tgggcacttc agttaggacc taaggatgcc agccggcagg tttatatatg cagcaacaat  720 attcaagcgc gacaacaggt tattgaactt gcccgccagt tgaatttcat tcccattgac  780 ttgggatcct tatcatcagc cagagagatt gaaaatttac ccctacgact ctttactctc  840 tggagagggc cagtggtggt agctataagc ttggccacat tttttttcct ttattccttt  900 gtcagagatg tgattcatcc atatgctaga aaccaacaga gtgactttta caaaattcct  960 atagagattg tgaataaaac cttacctata gttgccatta ctttgctctc cctagtatac 1020 cttgcaggtc ttctggcagc tgcttatcaa ctttattacg gcaccaagta taggagattt 1080 ccaccttggt tggaaacctg gttacagtgt agaaaacagc ttggattact aagttttttc 1140 ttcgctatgg tccatgttgc ctacagcctc tgcttaccga tgagaaggtc agagagatat 1200 ttgtttctca acatggctta tcagcaggtt catgcaaata ttgaaaactc ttggaatgag 1260 gaagaagttt ggagaattga aatgtatatc tcctttggca taatgagcct tggcttactt 1320 tccctcctgg cagtcacttc tatcccttca gtgagcaatg ctttaaactg gagagaattc 1380 agttttattc agtctacact tggatatgtc gctctgctca taagtacttt ccatgtttta 1440 atttatggat ggaaacgagc ttttgaggaa gagtactaca gattttatac accaccaaac 1500 tttgttcttg ctcttgtttt gccctcaatt gtaattctgg atcttttgca gctttgcaga 1560 tacccagact gagctggaac tggaatttgt cttcctattg actctacttc tttaaaagcg 1620 gctgcccatt acattcctca gctgtccttg cagttaggtg tacatgtgac tgagtgttgg 1680 ccagtgagat gaagtctcct caaaggaagg cagcatgtgt cctttttcat cccttcatct 1740 tgctgctggg attgtggata taacaggagc cctggcagct gtctccagag gatcaaagcc 1800 acacccaaag agtaaggcag attagagacc agaaagacct tgactacttc cctacttcca 1860 ctgctttttc ctgcatttaa gccattgtaa atctgggtgt gttacatgaa gtgaaaatta 1920 attctttctg cccttcagtt ctttatcctg ataccattta acactgtctg aattaactag 1980 actgcaataa ttctttcttt tgaaagcttt taaaggataa tgtgcaattc acattaaaat 2040 tgattttcca ttgtcaatta gttatactca ttttcctgcc ttgatctttc attagatatt 2100 ttgtatctgc ttggaatata ttatcttctt tttaactgtg taattggtaa ttactaaaac 2160 tctgtaatct ccaaaatatt gctatcaaat tacacaccat gttttctatc attctcatag 2220 atctgcctta taaacattta aataaaaagt actatttaat gatttaactt ctgttttgaa 2280 aaaaaaaaaa aaaaaaaaaa

TABLE LIII(b) Nucleotide sequence alignment of 98P4B6 v.1 (SEQ ID NO: 158) and 98P4B6 v.3 (SEQ ID NO: 159) Score = 4013 bits (2087), Expect = 0.0 Identities = 2116/2128 (99%), Gaps = 1/2128 (0%) Strand = Plus/Plus

NOTE: AN INSERTION OF A SINGLE BASE AT 1845 OF V.3

TABLE LV(b) Peptide sequences of protein coded by 98P4B6 v.3 (SEQ ID NO: 160) MESISMMGSP KSLSETCLPN GINGIKDARK VTVGVIGSGD  60 FAKSLTIRLI RCGYHVVIGS RNPKFASEFF PHVVDVTHHE DALTKTNIIF VAIHREHYTS 120 LWDLRHLLVG KILIDVSNNM RINQYPESNA EYLASLFPDS LIVKGFNVVS AWALQLGPKD 180 ASRQVYICSN NIQARQQVIE IARQLNFIPI DLGSLSSARE IENLPLRLFT LWRGPVVVAI 240 SLATFFFLYS FVRDVIHPYA RNQQSDFYKI PIEIVNKTLP IVAITLLSLV YLAGLLAAAY 300 QLYYGTKYRR FPPWLETWLQ CRKQLGLLSF FFAMVHVAYS LCLPMRRSER YLFLNMAYQQ 360 VHANIENSWN EEEVWRIEMY ISFGIMSLGL LSLLAVTSIP SVSNALNWRE FSFIQSTLGY 420 VALLISTFHV LIYGWKRAFE EEYYRFYTPP NFVLALVLPS IVILDLLQLC RYPD 454

TABLE LV(b) Amino acid sequence alignment of 98P4B6v.1 (SEQ ID NO: 161) and 98P4B6 v.3 (SEQ ID NO: 162) Score = 910 bits (2351), Expect = 0.0 Identities = 454/454 (100%), Positives = 454/454 (100%) V.1: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS V.3: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 V.1: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 V.3: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 V.1: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE V.3: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 V.1: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA V.3: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 V.1: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ V.3: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 V.1: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY V.3: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 V.1: 361 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE 420 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE V.3: 361 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE 420 V.1: 421 EEYYRFYTPPNFVLALVLPSIVILDLLQLCRYPD 454 EEYYRFYTPPNFVLALVLPSIVILDLLQLCRYPD V.3: 421 EEYYRFYTPPNFVLALVLPSIVILDLLQLCRYPD 454

TABLE LII(c) Nucleotide sequence of transcript variant 98P4B6 v.4 (SEQ ID NO: 163) cccacgcgtc cgcggacgcg tgggcggacg cgtgggttcc   60 tcgggccctc ggcgccacaa gctgtccggg cacgcagccc ctagcggcgc gtcgctgcca  120 agccggcctc cgcgcgcctc cctccttcct tctcccctgg ctgttcgcga tccagcttgg  180 gtaggcgggg aagcagctgg agtgcgaccg ccacggcagc caccctgcaa ccgccagtcg  240 gagagctaag ggcaagtcct gaggttgggc ccaggagaaa gaaggcaagg agacattgtc  300 ccaggatatt cttggtgatc ttggaagtgt ccgtatcatg gaatcaatct ctatgatggg  360 aagccctaag agccttagtg aaacttgttt acctaatggc ataaatggta tcaaagatgc  420 aaggaaggtc actgtaggtg tgattggaag tggagatttt gccaaatcct tgaccattcg  480 acttattaga tgcggctatc atgtggtcat aggaagtaga aatcctaagt ttgcttctga  540 attttttcct catgtggtag atgtcactca tcatgaagat gctctcacaa aaacaaatat  600 aatatttgtt gctatacaca gagaacatta tacctccctg tgggacctga gacatctgct  660 tgtgggtaaa atcctgattg atgtgagcaa taacatgagg ataaaccagt acccagaatc  720 caatgctgaa tatttggctt cattattccc agattctttg attgtcaaag gatttaatgt  780 tgtctcagct tgggcacttc agttaggacc taaggatgcc agccggcagg tttatatatg  840 cagcaacaat attcaagcgc gacaacaggt tattgaactt gcccgccagt tgaatttcat  900 tcccattgac ttgggatcct tatcatcagc cagagagatt gaaaatttac ccctacgact  960 ctttactctc tggagagggc cagtggtggt agctataagc ttggccacat tttttttcct 1020 ttattccttt gtcagagatg tgattcatcc atatgctaga aaccaacaga gtgactttta 1080 caaaattcct atagagattg tgaataaaac cttacctata gttgccatta ctttgctctc 1140 cctagtatac cttgcaggtc ttctggcagc tgcttatcaa ctttattacg gcaccaagta 1200 taggagattt ccaccttggt tggaaacctg gttacagtgt agaaaacagc ttggattact 1260 aagttttttc ttcgctatgg tccatgttgc ctacagcctc tgcttaccga tgagaaggtc 1320 agagagatat ttgtttctca acatggctta tcagcaggtt catgcaaata ttgaaaactc 1380 ttggaatgag gaagaagttt ggagaattga aatgtatatc tcctttggca taatgagcct 1440 tggcttactt tccctcctgg cagtcacttc tatcccttca gtgagcaatg ctttaaactg 1500 gagagaattc agttttattc agtctacact tggatatgtc gctctgctca taagtacttt 1560 ccatgtttta atttatggat ggaaacgagc ttttgaggaa gagtactaca gattttatac 1620 accaccaaac tttgttcttg ctcttgtttt gccctcaatt gtaattctgg atcttttgca 1680 gctttgcaga tacccagact gagctggaac tggaatttgt cttcctattg actctacttc 1740 tttaaaagcg gctgcccatt acattcctca gctgtccttg cagttaggtg tacatgtgac 1800 tgagtgttgg ccagtgagat gaagtctcct caaaggaagg cagcatgtgt cctttttcat 1860 cccttcatct tgctgctggg attgtggata taacaggagc cctggcagct gtctccagag 1920 gatcaaagcc acacccaaag agtaaggcag attagagacc agaaagacct tgactacttc 1980 cctacttcca ctgcttttcc tgcatttaag ccattgtaaa tctgggtgtg ttacatgaag 2040 tgaaaattaa ttctttctgc ccttcagttc tttatcctga taccatttaa cactgtctga 2100 attaactaga ctgcaataat tctttctttt gaaagctttt aaaggataat gtgcaattca 2160 cattaaaatt gattttccat tgtcaattag ttatactcat tttcctgcct tgatctttca 2220 ttagatattt tgtatctgct tggaatatat tatcttcttt ttaactgtgt aattggtaat 2280 tactaaaact ctgtaatctc caaaatattg ctatcaaatt acacaccatg ttttctatca 2340 ttctcataga tctgccttat aaacatttaa ataaaaagta ctatttaatg attt 2374

TABLE LIII(c) Nucleotide sequence alignment of 98P4B6 v.1 (SEQ ID NO: 164) and 98P4B6 v.4 (SEQ ID NO: 165) Score = 404 bits (210), Expect = e-109 Identities = 210/210 (100%) Strand = Plus/Plus

Score = 4022 bits (2092), Expect = 0.0 Identities = 2092/2092 (100%) Strand = Plus/Plus

TABLE LIV(c) Peptide sequences of protein coded by 98P4B6 v.4 (SEQ ID NO: 166) MESISMMGSP KSLSETCLPN GINGIKDARK VTVGVIGSGD  60 FAKSLTIRLI RCGYHVVIGS RNPKFASEFF PHVVDVTHHE DALTKTNIIF VAIHREHYTS 120 LWDLRHLLVG KILIDVSNNM RINQYPESNA EYLASLFPDS LIVKGFNVVS AWALQLGPKD 180 ASRQVYICSN NIQARQQVIE LARQLNFIPI DLGSLSSARE IENLPLRLFT LWRGPVVVAI 240 SLATFFFLYS FVRDVIHPYA RNQQSDFYKI PTEIVNKTLP IVAITLLSLV YLAGLLAAAY 300 QLYYGTKYRR FPPWLETWLQ CRKQLGLLSF FFANVHVAYS LCLPMRRSER YLFLNMAYQQ 360 VHANTENSWN EEEVWRIEMY ISFGIMSLGL LSLLAVTSIP SVSNALNWRE FSFIQSTLGY 420 VALLISTFHV LIYGWKRAFE EEYYRFYTPP NFVLALVLPS IVILDLLQLC RYPD 454

TABLE LV(c) Amino acid sequence alignment of 98P4B6 v.1 (SEQ ID NO: 167) and 98P4B6 v.4 (SEQ ID NO: 168) Score = 910 bits (2351), Expect = 0.0 Identities = 454/454 (100%), Positives = 454/454 (100%) V.1: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS V.4: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 V.1: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM V.4: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 V.1: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE V.4: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 V.1: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA V.4: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 V.1: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ V.4: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 V.1: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY V.4: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 V.1: 361 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE 420 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE V.4: 361 ISFGIMSLGLLSLLAVTSTPSVSNAIMWREFSFIQSTLGYVAILISTFHVLIYGWKRAFE 420 V.1: 421 EEYYRFYTPPNFVLALVLPSIVILDLLQLCRYPD 454 EEYYRFYTPPNFVLAIVLPSIVILDLLQLCRYPD V.4: 421 EEYYRFYTPPNFVLALVLPSIVILDLLQLCRYPD 454

TABLE LII(d) Nucleotide sequence of transcript variant 98P4B6 v.5 (SEQ ID NO: 169) cccacgcgtc cgcggacgcg tgggcggacg cgtgggttcc   60 tcgggccctc ggcgccacaa gctgtccggg cacgcagccc ctagcggcgc gtcgctgcca  120 agccggcctc cgcgcgcctc cctccttcct tctcccctgg ctgttcgcga tccagcttgg  180 gtaggcgggg aagcagctgg agtgcgaccg ctacggcagc caccctgcaa ccgccagtcg  240 gagagctaag ggcaagtcct gaggttgggc ccaggagaaa gaaggcaagg agacattgtc  300 ccaggatatt cttggtgatc ttggaagtgt ccgtatcatg gaatcaatct ctatgatggg  360 aagccctaag agccttagtg aaacttgttt acctaatggc ataaatggta tcaaagatgc  420 aaggaaggtc actgtaggtg tgattggaag tggagatttt gccaaatcct tgaccattcg  480 acttattaga tgcggctatc atgtggtcat aggaagtaga aatcctaagt ttgcttctga  540 attttttcct catgtggtag atgtcactca tcatgaagat gctctcacaa aaacaaatat  600 aatatttgtt gctatacaca gagaacatta tacctccctg tgggacctga gacatctgct  660 tgtgggtaaa atcctgattg atgtgagcaa taacatgagg ataaaccagt acccagaatc  720 caatgctgaa tatttggctt cattattccc agattctttg attgtcaaag gatttaatgt  780 tgtctcagct tgggcacttc agttaggacc taaggatgcc agccggcagg tttatatatg  840 cagcaacaat attcaagcgc gacaacaggt tattgaactt gcccgccagt tgaatttcat  900 tcccattgac ttgggatcct tatcatcagc cagagagatt gaaaatttac ccctacgact  960 ctttactttc tggagagggc cagtggtggt agctataagc ttggccacat tttttttcct 1020 ttattccttt gtcagagatg tgattcatcc atatgctaga aaccaacaga gtgactttta 1080 caaaattcct atagagattg tgaataaaac cttacctata gttgccatta ctttgctctc 1140 cctagtatac cttgcaggtc ttctggcagc tgcttatcaa ctttattacg gcaccaagta 1200 taggagattt ccaccttggt tggaaacctg gttacagtgt agaaaacagc ttggattact 1260 aagttttttc ttcgctatgg tccatgttgc ctacagcctc tgcttaccga tgagaaggtc 1320 agagagatat ttgtttctca acatggctta tcagcaggtt catgcaaata ttgaaaactc 1380 ttggaatgag gaagaagttt ggagaattga aatgtatatc tcctttggca taatgagcct 1440 tggcttactt tccctcctgg cagtcacttc tatcccttcg gtgagcaatg ctttaaactg 1500 gagagaattc agttttattc agatcttttg cagctttgca gatacccaga ctgagctgga 1560 actggaattt gtcttcctat tgactctact tctttaaaag cggctgccca ttacattcct 1620 cagctgtcct tgcagttagg tgtacatgtg actgagtgtt ggccagtgag atgaagtctc 1680 ctcaaaggaa ggcagcatgt gtcctttttc atcccttcat cttgctgctg ggattgtgga 1740 tataacagga gccctggcag ctgctccaga ggatcaaagc cacacccaaa gagtaaggca 1800 gattagagac cagaaagacc ttgactactt ccctacttcc actgcttttt cctgcattta 1860 agccattgta aatctgggtg tgttacatga agtgaaaatt aattctttct gcccttcagt 1920 tctttatcct gataccattt aacactgtct gaattaacta gactgcaata attctttctt 1980 ttgaaagctt ttaaaggata atgtgcaatt cacattaaaa ttgattttcc attgtcaatt 2040 agttatactc attttcctgc cttgatcttt cattagatat tttgtatctg cttggaatat 2100 attatcttct ttttaactgt gtaattggta attactaaaa ctctgtaatc tccaaaatat 2160 tgctatcaaa ttacacacca tgttttctat cattctcata gatctgcctt ataaacattt 2220 aaataaaaag tactatttac caaaaaaaaa aaaaaaaaaa aaaaaaaaa 2249

TABLE LIII(d) Nucleotide sequence alignment of 98P4B6 v.1 (SEQ ID NO: 170) and 98P4B6 v.5 (SEQ ID NO: 171) Score = 398 bits (207), Expect = e-107 Identities = 209/210 (99%) Strand = Plus/Plus

Score = 2334 bits (1214), Expect = 0.0 Identities = 1218/1220 (99%) Strand = Plus/Plus

Score = 1375 bits (715), Expect = 0.0 Identities = 741/749 (98%), Gaps = 2/749 (0%) Strand = Plus/Plus

NOTE: A SNP AT 192 AND AT 1510, A DELETION AT 1742-1743, AND AN INSERTION OF SINGLE BASE AT 1830 OF V.5.

TABLE LIV(d) Peptide sequences of protein coded by 98P4B6 v.5 (SEQ ID NO: 172) MESISMMGSP KSLSETCLPN GINGIKDARK VTVGVIGSGD  60 FAKSLTIRLI RCGYHVVIGS RNPKFASEFF PHVVDVTHHE DALTKTNIIF VAIHREHYTS 120 LWDLRHLLVG KILIDVSNNM RINQYPESNA EYLASLFPDS LIVKGFNVVS AWALQLGPKD 180 ASRQVYICSN NIQARQQVIE LARQLNFIPI DLGSLSSARE IENLPLRLFT FWRGPVVVAI 240 SLATFFFLYS FVRDVIHPYA RNQQSDFYKI PIEIVNKTLP IVAITLLSLV YLAGLLAAAY 300 QLYYGTKYRR FPPWLETWLQ CRKQLGLLSF FFAMVHVAYS LCLPMRRSER YLFLNMAYQQ 360 VHANIENSWN EEEVWRIEMY ISFGIMSLGL LSLLAVTSIP SVSNALNWRE FSFIQIFCSF 419 ADTQTELELE FVFLLTLLL

TABLE LV(d) Amino acid sequence alignment of 98P4B6 v.1 (SEQ ID NO: 173) and 98P4B6 v.5 (SEQ ID NO: 174) Score = 788 bits (2036), Expect = 0.0 Identities = 394/395 (99%), Positives = 394/395 (99%) V.1: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS V.5: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 V.1: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM V.5: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 V.1: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE V.5: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 V.1: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTFWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 LARQLNFIPIDLGSLSSAREIENLPLRLFT WRGPVVVAISLATFFFLYSFVRDVIHPYA V.5: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTFWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 V.1: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ V.5: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 V.1: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY V.5: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 V.1: 361 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQ 395 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQ V.5: 361 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQ 395 NOTE: A SNP CAUSED A SINGEL AMINO ACID DIFFERENCE AT 211.

TABLE LII(e) Nucleotide sequence of transcnpt variant 98P4B6 v.6 (SEQ ID NO: 175) cccacgcgtc cgcggacgcg tgggcggacg cgtgggttcc   60 tcgggccctc ggcgccacaa gctgtccggg cacgcagccc ctagcggcgc gtcgctgcca  120 agccggcctc cgcgcgcctc cctccttcct tctcccctgg ctgttcgcga tccagcttgg  180 gtaggcgggg aagcagctgg agtgcgaccg ccacggcagc caccctgcaa ccgccagtcg  240 gagagctaag ggcaagtcct gaggttgggc ccaggagaaa gaaggcaagg agacattgtc  300 ccaggatatt cttggtgatc ttggaagtgt ccgtatcatg gaatcaatct ctatgatggg  360 aagccctaag agccttagtg aaacttgttt acctaatggc ataaatggta tcaaagatgc  420 aaggaaggtc actgtaggtg tgattggaag tggagatttt gccaaatcct tgaccattcg  480 acttattaga tgcggctatc atgtggtcat aggaagtaga aatcctaagt ttgcttctga  540 attttttcct catgtggtag atgtcactca tcatgaagat gctctcacaa aaacaaatat  600 aatatttgtt gctatacaca gagaacatta tacctccctg tgggacctga gacatctgct  660 tgtgggtaaa atcctgattg atgtgagcaa taacatgagg ataaaccagt acccagaatc  720 caatgctgaa tatttggctt cattattccc agattctttg attgtcaaag gatttaatgt  780 tgtctcagct tgggcacttc agttaggacc taaggatgcc agccggcagg tttatatatg  840 cagcaacaat attcaagcgc gacaacaggt tattgaactt gcccgccagt tgaatttcat  900 tcccattgac ttgggatcct tatcatcagc cagagagatt gaaaatttac ccctacgact  960 ctttactctc tggagagggc cagtggtggt agctataagc ttggccacat tttttttcct 1020 ttattccttt gtcagagatg tgattcatcc atatgctaga aaccaacaga gtgactttta 1080 caaaattcct atagagattg tgaataaaac cttacctata gttgccatta ctttgctctc 1140 cctagtatac cttgcaggtc ttctggcagc tgcttatcaa ctttattacg gcaccaagta 1200 taggagattt ccaccttggt tggaaacctg gttacagtgt agaaaacagc ttggattact 1260 aagttttttc ttcgctatgg tccatgttgc ctacagcctc tgcttaccga tgagaaggtc 1320 agagagatat ttgtttctca acatggctta tcagcaggtt catgcaaata ttgaaaactc 1380 ttggaatgag gaagaagttt ggagaattga aatgtatatc tcctttggca taatgagcct 1440 tggcttactt tccctcctgg cagtcacttc tatcccttca gtgagcaatg ctttaaactg 1500 gagagaattc agttttattc agtctacact tggatatgtc gctctgctca taagtacttt 1560 ccatgtttta atttatggat ggaaacgagc ttttgaggaa gagtactaca gattttatac 1620 accaccaaac tttgttcttg ctcttgtttt gccctcaatt gtaattctgg gtaagattat 1680 tttattcctt ccatgtataa gccgaaagct aaaacgaatt aaaaaaggct gggaaaagag 1740 ccaatttctg gaagaaggta ttggaggaac aattcctcat gtctccccgg agagggtcac 1800 agtaatgtga tgataaatgg tgttcacagc tgccatataa agttctactc atgccattat 1860 ttttatgact tctacgttca gttacaagta tgctgtcaaa ttatcgtggg ttgaaacttg 1920 ttaaatgaga tttcaactga cttagtgata gagttttctt caagttaatt ttcacaaatg 1980 tcatgtttgc caatatgaat ttttctagtc aacatattat tgtaatttag gtatgttttg 2040 ttttgttttg cacaactgta accctgttgt tactttatat ttcataatca gacaaaaata 2100 cttacagtta ataatataga tataatgtta aaaacaattt gcaaaccagc agaattttaa 2160 gcttttaaaa taattcaatg gatatacatt tttttctgaa gattaagatt ttaattattc 2220 aacttaaaaa gtagaaatgc attattatac atttttttaa gaaaggacac gttatgttag 2280 catctaggta aggctgcatg atagcattcc tatatttctc tcataaaata ggatttgaag 2340 gatgaaatta attgtatgaa gcaatgtgat tatatgaaga gacacaaatt aaaaagacaa 2400 attaaacctg aaattatatt taaaatatat ttgagacatg aaatacatac tgataataca 2460 tacctcatga aagattttat tctttattgt gttacagagc agtttcattt tcatattaat 2520 atactgatca ggaagaggat tcagtaacat ttggcttcca aaactgctat ctctaatacg 2580 gtaccaatcc taggaactgt atactagttc ctacttagaa caaaagtatc aagtttgcac 2640 acaagtaatc tgccagctga cctttgtcgc accttaacca gtcaccactt gctatggtat 2700 aggattatac tgatgttctt tgagggattc tgatgtgcta ggcatggttc taagtacttt 2760 acttgtatta tcccatttaa tacttagaac aaccccgtga gataagtagt tattatcctc 2820 attttacaca tgagggaccg aaggatagaa aagttatttt tcaaaggtct tgcagttaat 2880 aaatggcaga gtgagcattc aagtccaggt agtcatattc cagaggccac ggttttaacc 2940 actaggctct agagctcccg ccgcgcccct atgcattatg ttcacaatgc caatctagat 3000 gcttcctctt ttgtataaag tcactgacat tctttagagt gggttgggtg catccaaaaa 3060 tgtataaaaa tattattata ataaacttat tactgcttgt agggtaattc acagttactt 3120 accctattct tgcttggaac atgagcctgg agacccatgg cagtccatat gcctccctat 3180 gcagtgaagg gccctagcag tgttaacaaa ttgctgagat cccacggagt ctttcaaaaa 3240 tctctgtaga gttagtcttc tccttttctc ttcctgagaa gttctcctgc ctgcataacc 3300 attcattagg gagtacttta caagcatgaa ggatattagg gtaagtggct aattataaat 3360 ctactctaga gacatataat catacagatt attcataaaa tttttcagtg ctgtccttcc 3420 acatttaatt gcattttgct caaactgtag aatgccctac attcccccca ccccaatttg 3480 ctatttcctt attaaaatag aaaattatag gcaagataca attatatgcg ttcctcttcc 3540 tgaaattata acatttctaa acttacccac gtagggacta ctgaatccaa ctgccaacaa 3600 taaaaagact tttatttagt agaggctacc tttcccccca gtgactcttt ttctacaact 3660 gccttgtcag tttggtaatt cacttatgat tttctaatgt tctcttggtg aattttatta 3720 tcttggaccc tctttttttt tttttttaaa gacagagtct tgctctgtca ccca 3754

TABLE LIII(e) Nucleotide sequence alignment off 98P4B6 v.1 (SEQ ID NO: 176) and 98P4B6 v.6 (SEQ ID NO: 177) Score = 404 bits (210), Expect = e-109 Indentities = 210/210 (100%) Strand = Plus/Plus

Score = 2630 bits (1368), Expect = 0.0 Identities = 1368/1368 (100%) Strand = Plus/Plus

TABLE LIV(e) Peptide sequences of protein coded by 98P4B6 v.6 (SEQ ID NO: 178) MESISMMGSP KSLSETCLPN GINGIKDARK VTVGVIGSGD  60 FAKSLTIRLI RCGYHVVIGS RNPKFASEFF PHVVDVTHHE DALTKTNIIF VAIHREHYTS 120 LWDLRHLLVG KILIDVSNNM RINQYPESNA EYLASLFPDS LIVKGFNVVS AWALQLGPKD 180 ASRQVYICSN NIQARQQVIE LARQLNFIPI DLGSLSSARE IENLPLRLFT LWRGPVVVAI 240 SLATFFFLYS FVRDVIHPYA RNQQSDFYKI PIEIVNKTLP IVAITLLSLV YLAGLLAAAY 300 QLYYGTKYRR FPPWLETWLQ CRKQLGLLSF FFAMVHVAYS LCLPMRRSER YLFLNMAYQQ 360 VHANIENSWN EEEVWRIEMY ISFGIMSLGL LSLLAVTSIP SVSNALNWRE FSFIQSTLGY 420 VALLISTFHV LIYGWKRAFE EEYYRFYTPP NFVLALVLPS IVILGKIILF LPCISRKLKR 480 IKKGWEKSQF LEEGIGGTIP HVSPERVTVM 490

TABLE LV(e) Amino acid sequence alignment of 98P4B6 v.1 (SEQ ID NO: 179) and 98P4B6 v.6 (SEQ ID NO: 180) Score = 888 bits (2294), Expect = 0.0 Identities = 444/444 (100%), Positives = 444/444 (100%) V.1: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS V.6: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 V.1: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM V.6: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 V.1: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE V.6: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 V.1: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA V.6: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 V.1: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ V.6: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 V.1: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY V.6: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 V.1: 361 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE 420 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE V.6: 361 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE 420 V.1: 421 EEYYRFYTPPNFVLALVLPSIVIL 444 EEYYRFYTPPNFVLALVLPSIVIL V.6: 421 EEYYRFYTPPNFVLALVLPSIVIL 444

TABLE LII(f) Nucleotide sequence of transcript variant 98P4B6 v.7 (SEQ ID NO: 181) ggagaaaatt tacagaaacc cagagccaaa ggtgctctca   60 ggggatcccc tgaaacattc aaagccattg cggccccaga agcttgggta ggcggggaag  120 cagctggagt gcgaccgccg cggcagccac cctgcaaccg ccagtcggag gtgcagtccg  180 taggccctgg cccccgggtg ggcccttggg gagtcggcgc cgctcccggg gagctgcaag  240 gctcgcccct gcccggcgtg gagggcgcgg ggggcgcgga ggatattctt ggtgatcttg  300 gaagtgtccg tatcatggaa tcaatctcta tgatgggaag ccctaagagc cttagtgaaa  360 cttttttacc taatggcata aatggtatca aagatgcaag gaaggtcact gtaggtgtga  420 ttggaagtgg agattttgcc aaatccttga ccattcgact tattagatgc ggctatcatg  480 tggtcatagg aagtagaaat cctaagtttg cttctgaatt ttttcctcat gtggtagatg  540 tcactcatca tgaagatgct ctcacaaaaa caaatataat atttgttgct atacacagag  600 aacattatac ctccctgtgg gacctgagac atctgcttgt gggtaaaatc ctgattgatg  660 tgagcaataa catgaggata aaccagtacc cagaatccaa tgctgaatat ttggcttcat  720 tattcccaga ttctttgatt gtcaaaggat ttaatgttgt ctcagcttgg gcacttcagt  780 taggacctaa ggatgccagc cggcaggttt atatatgcag caacaatatt caagcgcgac  840 aacaggttat tgaacttgcc cgccagttga atttcattcc cattgacttg ggatccttat  900 catcagccag agagattgaa aatttacccc tacgactctt tactctctgg agagggccag  960 tggtggtagc tataagcttg gccacatttt ttttccttta ttcctttgtc agagatgtga 1020 ttcatccata tgctagaaac caacagagtg acttttacaa aattcctata gagattgtga 1080 ataaaacctt acctatagtt gccattactt tgctctccct agtatacctc gcaggtcttc 1140 tggcagctgc ttatcaactt tattacggca ccaagtatag gagatttcca ccttggttgg 1200 aaacctggtt acagtgtaga aaacagcttg gattactaag ttttttcttc gctatggtcc 1260 atgttgccta cagcctctgc ttaccgatga gaaggtcaga gagatatttg tttctcaaca 1320 tggcttatca gcagtctaca cttggatatg tcgctctgct cataagtact ttccatgttt 1380 taatttatgg atggaaacga gcttttgagg aagagtacta cagattttat acaccaccaa 1440 actttgttct tgctcttgtt ttgccctcaa ttgtaattct ggatctgtct gtggaggttc 1500 tggcttcccc agctgctgcc tggaaatgct taggtgctaa tatcctgaga ggaggattgt 1560 cagagatagt actccccata gagtggcagc aggacaggaa gatcccccca ctctccaccc 1620 cgccgccacc ggccatgtgg acagaggaag ccggggcgac cgccgaggcc caggaatccg 1680 gcatcaggaa caagtctagc agttccagtc aaatcccggt ggttggggtg gtgacggagg 1740 acgatgaggc gcaggattcc attgatcccc cagagagccc tgatcgtgcc ttaaaagccg 1800 cgaattcctg gaggaaccct gtcctgcctc acactaatgg tgtggggcca ctgtgggaat 1860 tcctgttgag gcttctcaaa tctcaggctg cgtcaggaac cctgtctctt gcgttcacat 1920 cctggagcct tggagagttc cttgggagtg ggacatggat gaagctggaa accataattc 1980 tcagcaaact aacacaggaa cagaaatcca aacactgcat gttctcactg ataagtggga 2040 gttgaacaat gagaacacat ggacacaggg aggggaacgt cacacaccag ggcctgtcgg 2100 gggtgggagg cctagcaatt cattagaatt acctgtgaag cttttaaaat gtaaggtttg 2160 gatggaatgc tcagacccta ccttagaccc aattaagccc acagctttga gg 2192

TABLE LIII(f) Nucleotide sequence alignment of 98P4B6 v.1 (SEQ ID NO: 182) and 98P4B6 v.7 (SEQ ID NO: 183) Score = 2350 bits (1222), Expect = 0.0 Identities = 1230/1234 (99%) Strand = Plus/Plus

Score = 298 bits (155), Expect = 2e-77 Identities = 155/155 (100%) Strand = Plus/Plus

TABLE LIV(f) Peptide sequences of protein coded by 98P4B6 v.7 (SEQ ID NO: 184) MESISMMGSP KSLSETFLPN GINGIKDARK VTVGVIGSGD  60 FAKSLTIRLI RCGYHVVIGS RNPKFASEFF PHVVDVTHHE DALTKTNIIF VAIHREHYTS 120 LWDLRHLLVG KILIDVSNNM RINQYPESNA EYLASLFPDS LIVKGFNVVS AWALQLGPKD 180 ASRQVYICSN NIQARQQVIE LARQLNFIPI DLGSLSSARE IENLPLRLFT LWRGPVVVAI 240 SLATFFFLYS FVRDVIHPYA RNQQSDFYKI PIEIVNKTLP IVAITLLSLV YLAGLLAAAY 300 QLYYGTKYRR FPPWLETWLQ CRKQLGLLSF FFAMVHVAYS LCLPMRRSER YLFLNMAYQQ 360 STLGYVALLI STFHVLIYGW KRAFEEEYYR FYTPPNFVLA LVLPSIVILD LSVEVLASPA 420 AAWKCLGANI LRGGLSEIVL PIEWQQDRKI PPLSTPPPPA MWTEEAGATA EAQESGIRNK 480 SSSSSQIPVV GVVTEDDEAQ DSIDPPESPD RALKAANSWR NPVLPHTNGV GPLWEFLLRL 540 LKSQAASGTL SLAFTSWSLG EFLGSGTWMK LETIILSKLT QEQKSKHCMF SLISGS 576

TABLE LV(f) Amino acid sequence alignment of 98P4B6 v.1 (SEQ ID NO: 185) and 98P4B6 v.7 (SEQ ID NO: 186) Score = 753 bits (1944), Expect = 0.0 Identities = 390/446 (87%), Positives 390/446 (87%), Gaps = 55/446 (12%) V.1: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 MESISMMGSPKSLSET LPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS V.7: 1 MESISMMGSPKSLSETFLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 V.1: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM V.7: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 V.1: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE V.7: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 V.1: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA V.7: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 V.1: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ V.7: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 V.1: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQ V.7: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQ-------------------- 340 V.1: 361 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE 420                                    STLGYVALLISTFHVLIYGWKRAFE V.7: 341 -----------------------------------STLGYVALLISTFHVLIYGWKRAFE 365 V.1: 421 EEYYRFYTPPNFVLALVLPSIVILDL 446 EEYYRFYTPPNFVLALVLPSIVILDL V.7: 366 EEYYRFYTPPNFVLALVLPSIVILDL 391

TABLE LII(g) Nucleotide sequence of transcript variant 98P4B6 v.8 (SEQ ID NO: 187) gccccctccg agctccccga ctcctccccg cgctccacgg   60 ctcttcccga ctccagtcag cgttcctcgg gccctcggcg ccacaagctg tccgggcacg  120 cagcccctag cggcgcgtcg ctgccaagcc ggcctccgcg cgcctccctc cttccttctc  180 ccctggctgt tcgcgatcca gcttgggtag gcggggaagc agctggagtg cgaccgccac  240 ggcagccacc ctgcaaccgc cagtcggagg tgcagtccgt aggccctggc ccccgggtgg  300 gcccttgggg agtcggcgcc gctcccgagg agctgcaagg ctcgcccctg cccggcgtgg  360 agggcgcggg gggcgcggag gatattcttg gtgatcttgg aagtgtccgt atcatggaat  420 caatctctat gatgggaagc cctaagagcc ttagtgaaac ttgtttacct aatggcataa  480 atggtatcaa agatgcaagg aaggtcactg taggtgtgat tggaagtgga gattttgcca  540 aatccttgac cattcgactt attagatgcg gctatcatgt ggtcatagga agtagaaatc  600 ctaagtttgc ttctgaattt tttcctcatg tggtagatgt cactcatcat gaagatgctc  660 tcacaaaaac aaatataata tttgttgcta tacacagaga acattatacc tccctgtggg  720 acctgagaca tctgcttgtg ggtaaaatcc tgattgatgt gagcaataac atgaggataa  780 accagtaccc agaatccaat gctgaatatt tggcttcatt attcccagat tctttgattg  840 tcaaaggatt taatgttgtc tcagcttggg cacttcagtt aggacctaag gatgccagcc  900 ggcaggttta tatatgcagc aacaatattc aagcgcgaca acaggttatt gaacttgccc  960 gccagttgaa tttcattccc attgacttgg gatccttatc atcagccaga gagattgaaa 1020 atttacccct acgactcttt actctctgga gagggccagt ggtggtagct ataagcttgg 1080 ccacattttt tttcctttat tcctttgtca gagatgtgat tcatccatat gctagaaacc 1140 aacagagtga cttttacaaa attcctatag agattgtgaa taaaacctta cctatagttg 1200 ccattacttt gctctcccta gtataccttg caggtcttct ggcagctgct tatcaacttt 1260 attacggcac caagtatagg agatttccac cttggttgga aacctggtta cagtgtagaa 1320 aacagcttgg attactaagt tttttcttcg ctatggtcca tgttgcctac agcctctgct 1380 taccgatgag aaggtcagag agatatttgt ttctcaacat ggcttatcag caggttcatg 1440 caaatattga aaactcttgg aatgaggaag aagtttggag aattgaaatg tatatctcct 1500 ttggcataat gagccttggc ttactttccc tcctggcagt cacttctatc ccttcagtga 1560 gcaatgcttt aaactggaga gaattcagtt ttattcagtc tacacttgga tatgtcgctc 1620 tgctcataag tactttccat gttttaattt atggatggaa acgagctttt gaggaagagt 1680 actacagatt ttatacacca ccaaactttg ttcttgctct tgttttgccc tcaattgtaa 1740 ttctgggtaa gattatttta ttccttccat gtataagccg aaagctaaaa cgaattaaaa 1800 aaggctggga aaagagccaa tttctggaag aaggtatggg aggaacaatt cctcatgtct 1860 ccccggagag ggtcacagta atgtgatgac aaatggtgtt cacagctgcc atataaagtt 1920 ctactcatgc cattattttt atgacttcta cgttcagtta caagtatgct gtcaaattat 1980 cgtgggttga aacttgttaa atgagatttc aactgactta gtgatagagt tttcttcaag 2040 ttaattttca caaatgtcat gtttgccaat atgaattttt ctagtcaaca tattattgta 2100 atttaggtat gttttgtttt gttttgcaca actgtaaccc tgttgttact ttatatttca 2160 taatcaggca aaaatactta cagttaataa tatagatata atgttaaaaa caatttgcaa 2220 accagcagaa ttttaagctt ttaaaataat tcaatggata tacatttttt tctgaagatt 2280 aagattttaa ttattcaact taaaaagtag aaatgcatta ttatacattt ttttaagaaa 2340 ggacacgtta tgttagcatc taggtaaggc tgcatgatag cattcctata tttctctcat 2400 aaaataggat ttgaaggatg aaattaattg tatgaagcaa tgtgattata tgaagagaca 2460 caaattaaaa agacaaatta aacctgaaat tatatttaaa atatatttga gacatgaaat 2520 acatactgat aatacatacc tcatgaaaga ttttattctt tattgtgtta cagagcagtt 2580 tcattttcat attaatatac tgatcaggaa gaggattcag taacatttgg cttccaaaac 2640 tgctatctct aatacggtac caatcctagg aactgtatac tagttcctac ttagaacaaa 2700 agtatcaagt ttgcacacaa gtaatctgcc agctgacctt tgtcgcacct taaccagtca 2760 ccacttgcta tggtatagga ttatactgat gttctttgag ggattctgat gtgctaggca 2820 tggttctaag tactttactt gtattatccc atttaatact tagaacaacc ccgtgagata 2880 agtagttatt atcctcattt tacacatgag ggaccgaagg atagaaaagt tatttttcaa 2940 aggtcttgca gttaataaat ggcagagtga gcattcaagt ccaggtagtc atattccaga 3000 ggccacggtt ttaaccacta ggctctagag ctcccgccgc gcccctatgc attatgttca 3060 caatgccaat ctagatgctt cctcttttgt ataaagtcac tgacattctt tagagtgggt 3120 tgggtgcatc caaaaatgta taaaaatatt attataataa acttattact gcttgtaggg 3180 taattcacag ttacttaccc tattcttgct tggaacatga gcctggagac ccatggcagt 3240 ccatatgcct ccctatgcag tgaagggccc tagcagtgtt aacaaattgc tgagatccca 3300 cggagtcttt caaaaatctc tgtagagtta gtcttctcct tttctcttcc tgagaagttc 3360 tcctgcctgc ataaccattc attagggagt actttacaag catgaaggat attagggtaa 3420 gtggctaatt ataaatctac tctagagaca tataatcata cagattattc ataaaatttt 3480 tcagtgctgt ccttccacat ttaattgcat tttgctcaaa ctgtagaatg ccctacattc 3540 cccccacccc aatttgctat ttccttatta aaatagaaaa ttataggcaa gatacaatta 3600 tatgcgttcc tcttcctgaa attataacat ttctaaactt acccacgtag gtactactga 3660 atccaactgc caacaataaa aagactttta tttagtagag gctacctttc ccaccagtga 3720 ctctttttct acaactgcct tgtcagtttg gtaattcact tatgattttc taatgttctc 3780 ttggtgaatt ttattatctt gtaccctctt tttttttttt ttttttttta aagacagagt 3840 cttgctctgt cacccaggct ggagtgcagt ggcacgatct cggctcactg caagctctgc 3900 ctcccgggtt cacgccattc tcctgcctca gcctcccgag tagctgggac tacaggtgcc 3960 cgccaccatg cccggctgat ttctttttgt atttttagta gagacggagt ttcaccgtgt 4020 tagccaggat ggtctcgatc tcctgacctc gtgatccgcc cgccttggcc tccaaagtgc 4080 tgggattaca ggtgtgagct accgcgcccg gcctattatc ttgtactttc taactgagcc 4140 ctctattttc tttattttaa taatatttct ccccacttga gaatcacttg ttagttcttg 4200 gtaggaattc agttgggcaa tgataacttt tatgggcaaa aacattctat tatagtgaac 4260 taatgaaaat aacagcgtat tttcaatatt ttcttattcc ttaaattcca ctcttttaac 4320 actatgctta accacttaat gtgatgaaat attcctaaaa gttaaatgac tattaaagca 4380 tatattgttg catgtatata ttaagtagcc gatactctaa ataaaaatac cactgttaca 4440 gataaatggg gcctttaaaa atatgaaaaa caaacttgtg aaaatgtata aaagatgcat 4500 ctgttgtttc aaatggcact atcttctttt cagtactaca aaaacagaat aattttgaag 4560 ttttagaata aatgtaatat atttactata attctaaatg tttaaatgct tttctaaaaa 4620 tgcaaaacta tgatgtttag ttgctttatt ttacctctat gtgattattt ttcttaattg 4680 ttatttttta taatcattat ttttctgaac cattcttctg gcctcagaag taggactgaa 4740 ttctactatt gctaggtgtg agaaagtggt ggtgagaacc ttagagcagt ggagatttgc 4800 tacctggtct gtgttttgag aagtgcccct tagaaagtta aaagaatgta gaaaagatac 4860 tcagtcttaa tcctatgcaa aaaaaaaaat caagtaattg ttttcctatg aggaaaataa 4920 ccatgagctg tatcatgcta cttagctttt atgtaaatat ttcttatgtc tcctctatta 4980 agagtattta aaatcatatt taaatatgaa tctattcatg ctaacattat ttttcaaaac 5040 atacatggaa atttagccca gattgtctac atataaggtt tttatttgaa ttgtaaaata 5100 tttaaaagta tgaataaaat atatttatag gtatttatca gagatgatta ttttgtgcta 5160 catacaggtt ggctaatgag ctctagtgtt aaactacctg attaatttct tataaagcag 5220 cataaccttg gcttgattaa ggaattctac tttcaaaaat taatctgata atagtaacaa 5280 ggtatattat actttcatta caatcaaatt atagaaatta cttgtgtaaa agggcttcaa 5340 gaatatatcc aatttttaaa tattttaata tatctcctat ctgataactt aattcttcta 5400 aattaccact tgccattaag ctatttcata ataaattctg tacagtttcc ccccaaaaaa 5460 gagatttatt tatgaaatat ttaaagtttc taatgtggta ttttaaataa agtatcataa 5520 atgtaataag taaatattta tttaggaata ctgtgaacac tgaactaatt attcctgtgt 5580 cagtctatga aatccctgtt ttgaaatacg taaacagcct aaaatgtgtt gaaattattt 5640 tgtaaatcca tgacttaaaa caagatacat acatagtata acacacctca cagtgttaag 5700 atttatattg tgaaatgaga caccctacct tcaattgttc atcagtgggt aaaacaaatt 5760 ctgatgtaca ttcaggacaa atgattagcc ctaaatgaaa ctgtaataat ttcagtggaa 5820 actcaatctg tttttacctt taaacagtga attttacatg aatgaatggg ttcttcactt 5880 tttttttagt atgagaaaat tatacagtgc ttaattttca gagattcttt ccatatgtta 5940 ctaaaaaatg ttttgttcag cctaacatac tgagtttttt ttaactttct aaattattga 6000 atttccatca tgcattcatc caaaattaag gcagactgtt tggattcttc cagtggccag 6060 atgagctaaa ttaaatcaca aaagcagatg cttttgtatg atctccaaat tgccaacttt 6120 aaggaaatat tctcttgaaa ttgtctttaa agatcttttg cagctttgca gatacccaga 6180 ctgagctgga actggaattt gtcttcctat tgactctact tctttaaaag cggctgccca 6240 ttacattcct cagctgtcct tgcagttagg tgtacatgtg actgagtgtt ggccagtgag 6300 atgaagtctc ctcaaaggaa ggcagcatgt gtcctttttc atcccttcat cttgctgctg 6360 ggattgtgga tataacagga gccctggcag ctgtctccag aggatcaaag ccacacccaa 6420 agagtaaggc agattagaga ccagaaagac cttgactact tccctacttc cactgctttt 6480 tcctgcattt aagccattgt aaatctgggt gtgttacatg aagtgaaaat taattctttc 6540 tgcccttcag ttctttatcc tgataccatt taacactgtc tgaattaact agactgcaat 6600 aattctttct tttgaaagct tttaaaggat aatgtgcaat tcacattaaa attgattttc 6660 cattgtcaat tagttatact cattttcctg ccttgatctt tcattagata ttttgtatct 6720 gcttggaata tattatcttc tttttaactg tgtaattggt aattactaaa actctgtaat 6780 ctccaaaata ttgctatcaa attacacacc atgttttcta tcattctcat agatctgcct 6840 tataaacatt taaataaaaa gtactattta atgattt 6857

TABLE LIII(g) Nucleotide alignment of 98P4B6 v.1 (SEQ ID NO: 188) and 98P4B6 v.8 (SEQ ID NO: 189) Score = 3201 bits (1665), Expect = 0.0 Identities = 1665/1665 (100%) Strand = Plus/Plus

Score = 1381 bits (718), Expect = 0.0 Identities = 725/726 (99%), Gaps = 1/726 (0%) Strand = Plus/Plus

TABLE LIV(g) Peptide sequences of protein coded by 98P4B6 v.8 (SEQ ID NO: 190) MESISMMGSP KSLSETCLPN GINGIKDARK VTVGVIGSGD  60 FAKSLTIRLI RCGYHVVIGS RNPKFASEFF PHVVDVTHHE DALTKTNIIF VAIHREHYTS 120 LWDLRHLLVG KILIDVSNNM RINQYPESNA EYLASLFPDS LIVKGFNVVS AWALQLGPKD 180 ASRQVYICSN NIQARQQVIE LARQLNFIPI DLGSLSSARE IENLPLRLFT LWRGPVVVAI 240 SLATFFFLYS FVRDVIHPYA RNQQSDFYKI PIEIVNKTLP IVAITLLSLV YLAGLLAAAY 300 QLYYGTKYRR FPPWLETWLQ CRKQLGLLSF FFANVHVAYS LCLPMRRSER YLFLNMAYQQ 360 VHANTENSWN EEEVWRIEMY ISFGIMSLGL LSLLAVTSIP SVSNALNWRE FSFIQSTLGY 420 VALLISTFHV LIYGWKRAFE EEYYRFYTPP NFVLALVLPS IVILGKIILF LPCISRKLKR 480 IKKGWEKSQF LEEGMGGTIP HVSPERVTVM 490

TABLE LV(g) Amino acid sequence alignment of 98P4B6 v.1 (SEQ ID NO: 191) and 98P4B6 v.8 (SEQ ID NO: 192) Score = 888 bits (2294), Expect = 0.0 Identities = 444/444 (100%), Positives = 444/444 (100%) V.1: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS  60 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS V.8: 1 MESISMMGSPKSLSETCLPNGINGIKDARKVTVGVIGSGDFAKSLTIRLIRCGYHVVIGS 60 V.1: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM V.8: 61 RNPKFASEFFPHVVDVTHHEDALTKTNIIFVAIHREHYTSLWDLRHLLVGKILIDVSNNM 120 V.1: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE V.8: 121 RINQYPESNAEYLASLFPDSLIVKGFNVVSAWALQLGPKDASRQVYICSNNIQARQQVIE 180 V.1: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA V.8: 181 LARQLNFIPIDLGSLSSAREIENLPLRLFTLWRGPVVVAISLATFFFLYSFVRDVIHPYA 240 V.1: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ V.8: 241 RNQQSDFYKIPIEIVNKTLPIVAITLLSLVYLAGLLAAAYQLYYGTKYRRFPPWLETWLQ 300 V.1: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY V.8: 301 CRKQLGLLSFFFAMVHVAYSLCLPMRRSERYLFLNMAYQQVHANIENSWNEEEVWRIEMY 360 V.1: 361 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE 420 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE V.8: 361 ISFGIMSLGLLSLLAVTSIPSVSNALNWREFSFIQSTLGYVALLISTFHVLIYGWKRAFE 420 V.1: 421 EEYYRFYTPPNFVLALVLPSIVIL 444 EEYYRFYTPPNFVLALVLPSIVIL V.8: 421 EEYYRFYTPPNFVLALVLPSIVIL 444 

1. A method of inhibiting the growth of a paclitaxel-resistant prostate cancer cell that expresses a protein selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 27 and SEQ ID NO: 29, the method comprising exposing the cell to an antibody-agent conjugate comprising an antibody or fragment thereof conjugated to a cytotoxic agent or therapeutic agent, wherein the antibody or fragment thereof specifically binds to the protein.
 2. The method of claim 1 wherein the antibody or fragment is conjugated to a cytotoxic agent.
 3. The method of claim 2 wherein the cytotoxic agent is an auristatin or a maytansinoid.
 4. The method of claim 2 wherein the cytotoxic agent is a calcium channel inhibitor.
 5. The method of claim 4 wherein the calcium channel inhibitor is agatoxin. 